CN107913201A - The extract of microalgae and its application - Google Patents

Abstract

The present invention relates to a kind of cosmetic composition, it includes the extract of garlic Trentepohlia and/or Chaetoceros category and/or Chlorococcum microalgae and cosmetically acceptable auxiliary material, and the cosmetically acceptable auxiliary material is selected from C₁‑C₄Aliphatic alcohol, polyalcohol, oil ingredient, water and their mixture with 3 to 12 carbon atoms, wherein the extract is by using selected from C₁‑C₄Aliphatic alcohol, ethyl acetate, the solvent of water or their mixture handle the microalgae, and the extract of dissolving is removed from residue and is obtained from the pure extract of the solvent recovery.The extract especially shows excellent performance in terms of the metabolism of application on human skin and hair follicle is adjusted.The non-treatment method of human skin and/or hair is handled the invention further relates to the purposes of the cosmetic composition, and using it.

Description

The extract of microalgae and its application

It is that on October 13rd, 2011, Chinese Patent Application No. are 201180061149.5 (international application no the applying date that the application, which is, For PCT/EP2011/067945), the division Shen of the application for a patent for invention of entitled " extract of microalgae and its application " Please.

Technical field

The present invention relates to the field of cosmetics and bath article, and it is related to the extract of microalgae, obtains the side of the extract Method and composition, and their purposes in hair-care and skin care applications.

Background technology

Cosmetics and bath article industry particularly focus on anti-aging production looking for be suitable for preparing body care product A large amount of interest have had been put into the native compound of product.The reduction of the birthrate of population of industrialized economy body experience and service life Extension adds influence of the anti-aging product in health products and Cosmetic Market.

Wrinkle represents the obvious symptom of skin aging, and cosmetics are had been directed in this problem for many years.Dermal collagen Reduction and structural change be considered as wrinkle of skin produce the main reason for.In artificial synthesized compound and including micro- Collagen stimulant is actively screened in the natural component of algae extract.However, for for improving skin moisture-keeping and beauty Adjustment for the treatment of, it is also contemplated that epidermis.In epidermis, by nourishing top cellular layer, the undifferentiated keratinocyte of substrate Continuous propagation.These epidermal layer cells are moved by being divided into horn cell progressively towards skin surface.Horn cell is dead thin Born of the same parents, they form superficial cuticula, are removed eventually through decortication.The generation and loss of this continuous epidermal cell form skin It is constantly regenerating.Force to remove this regeneration of cuticula acceleration, and allow to the formation to anti-aging, papule and reduce scar Beauty influence.The treatment of some dermatopathies also may require that pressure and accelerate promoting epidermization.

However, in the major function of horny layer of epidermis, prevent water from undertaking very big importance from skin losses.Cuticula Thickness can not induce the illness of skin fragility and allergy completely, in the most severe case with xerosis and violent itch. Weighing apparatus stimulates keratinocyte differentiation to strengthen horny layer of epidermis, improves skin hydration effect, and be conducive to the healthy and smooth of skin.

On the one hand, some emerging economies, such as Brazil, strength contribution has been made for the growth of global Cosmetic Market, he Have very high demand in skin nursing, hair-care and perfume.It is the main of restriction U.S. that although anti-aging is thought in the Western countries Standard, but it is related to skin color in the identical standard of some Asian countries.Melanocyte is especially caused in skin and its attached The cell of melanocyte is generated in part.It must be taken into account that the potential instrumentality of melanin biosynthesis is used for cosmetics and medical applications In there are high interests.Skin-lightening cosmetic by response to Japan and other Asian countries in many people beauty it is expected, Meet the interest of more and more consumers.However, whitening product is also applied to treatment skin disease, such as, for example, yellowish-brown Spot, it is a kind of mainly to occur the skin disease of brown patch in cheekbone, forehead and upper lip.This problem has including Asian It is more common in the crowd of color skin.In client in the Western countries, whitening product is also appreciated, to prevent or inhibit Spot on the face, including brown spot and freckle, and the feature including anti-aging.

On the other hand, the compound is suitable for producing the positive regulator of melanocyte generation and obtains extensive cosmetics Using.Many people wish their tanned natural pale complexion and skin-color occur in the case where being not exposed to solar radiation It is plain calm.In addition, some catch at deeper and uniform hair color.For this reason, very safely effectively The dyeing brown stain agent of skin and hair is necessary.

In terms of this, it is assumed that the effective stimulant of melanin generation to trichochromes unit is especially relevant.Although Hair follicle has significant difference, but the natural adjusting of this biological function between neutralizing the metabolism of melanocyte in skin Discovery there are potential application in both organs.Prevent that hair from bleaching the target of the highly significant for representing cosmetics, at the same time It is related to beauty and anti-aging part.

Although lacking really effective solution, it is involved in the problems, such as hair follicle, mainly alopecia and pigmentation problems Treatment cause annual total market capitalisation more than 1,000,000,000 dollars.Alopecia represents the main problem to be solved, and 5- 5 alpha-reductases at present Inhibitor is considered as more active reagent.5- 5 alpha-reductases are to participate in the key enzyme that testosterone is converted into protona (DHT), It is considered as the major steroid compound for causing the alopecia in androgenetic alopecia.As commercially available minoxidil (falls strong (Rogaine)), the active production such as Finasteride (protect method and stop (Propecia)) and dutasteride (suitable urine is logical (Avodart)) Product should be administered in the case where doctor supervises, and cannot be used for treatment pregnant woman.Their curers to limited part Some side effects can be produced while giving gratifying reaction.Although advocate to promote the autonomic drug preparation of hair growth can be with Obtained with relatively low cost, but usually their effect is very limited amount of.

On the other hand, unnecessary hair also represent related cosmetics problem, and the novel non-toxic of hair growth inhibition The discovery of reagent can find relevant application.

Modern way of life, characterized by being frequently accompanied by the sedentary occupation of Nutrition Attitude of mistake, promotes in vivo extensively The excessive accumulation of fat.Many people are tormented by this problem, not only in their appearance and the relation of society, but also at them Health and life expectancy on have heavy consequence.In this respect, except stringent weight-reducing diet, exercise tired out or beautifying hand The dangerous and invasive intervening measure of art, few solutions.On the other hand, the people with normal type can be subject in spy The influence of fatty local deposits in the subcutaneous tissue of fixed body part.For example, cellulite is considered and this injustice The relevant typical problem of fat metabolism of weighing apparatus, it is scientifically defined as " lipodystrophy " or " oedema-collagenous fibres are hard Change ".Seldom cosmetic treatments can reduce subcutaneous layer of fat (also referred to as subcutaneous tissue) at present.Cosmetic industry is to being suitable for preventing The discovery of the compounds effective of lipolysis feels emerging very much in internal general fat accumulation and the subcutaneous tissue of promotion skin Interest.

In cosmetic field some examples, but the algae with considering here are provided using the relevant prior art of microalgae Species is relevant seldom.WO 1989/000606 is described to be used to produce Omega- using obligate and facultative marine eukaryotic microorganism 3 (n-3) aliphatic acid, it can be used for food, cosmetics, medical product.Except the specific Heterotrophic Culture of suggestion, which builds Purpose of the view for body treatment, the source of compound, particularly n-3 aliphatic acid is used as by the use of microalgae.Clearly propose Species are as valuable microalgae, such as diatom diamond shape algae (diatom Nitzschia) and hidden dinoflagellate (dinoflagellate Crypthecodinium cohni)。

Disclosed in nineteen ninety FR 2657012B1 (Secma) from Chlorophyceae, green branch algae guiding principle, Cryptophyceae, Bacillariophyceae (or silicon Algae) and general woods algae guiding principle obtain liquid extract Green Tea Extract activity.Due to GB 1392131A (Aubert etc.), from The utilization of Chaetoceros since 1975 for cosmetics is known.Japan Patent JP 3-822959B2 (Noevir KK) are related to And effectively preventing the skin lotion of wrinkle of skin, it includes the extract of specific diatom particularly Chaetoceros.Extraction solvent is selected from Ethanol, methanol, 1,3-BDO and water, and in the form of single or two or more mixed form use.Preferred Embodiment in, these solvents include inorganic salts and surfactant.US 5767095 (Photonz) discloses externally applied anti-inflammation Composition, among other things, it includes the single galactosyl-bis- eicosapentaenoics acylglycerol obtained from Chaetoceros and hailian seaweed (monogalactosyl-dieicosapentanoyl glycerol).It is green according to EP 1808483A1 (Cognis) lemon shape Ball algae (Chlorococcum citriforme) has been considered to the interesting source of the lutein of cosmetic applications.It is international special Profit application WO 1997/034489Al (aquaculture technology) are directed to use with the extract obtained from seaweed Chaetoceros or hailian seaweed As antibacterial activity agent, and it is related to comprising the composition for being used for the reagent for resisting pathogenic bacteria.International patent application WO 2010/0029115A1 (LVHM Recherche) proposes the purposes of some plant extracts, for example, obtained from hailian seaweed Extract is used to reduce skin and trichochromes is calm.

FR 2894473A1 (Daniel Jouvance) disclose from some microalgaes (Chromulina (Chromulina), Star bar algae (Asterionella) and four slit bamboo or chopped wood algaes (Tetraselmis)) obtain preparation purposes, to suppress aliphatic acid and lipid Metabolism in enzyme.Japan Patent JP 2000072642A1 (Lion) propose what is obtained from some kinds of macro Slimming preparation, however, the not available preparation on being obtained based on the microalgae strain considered from here carries out fat metabolism tune The prior art of section.

The prior art is kept silent completely on the problem of adjusting and lipolysis of keratoderma.Isochrysis galbana (Isochrysis) it is used for hair products with four slit bamboo or chopped wood algae extracts and is obtained respectively in EP2168570A2 and EP 2193785A2 To introduction.

The content of the invention

It is therefore an object of the present invention in order to reach the improved purpose in resistance and the decoloration of anti-alopecia-stopping and hair, open Hair, based on regenerative resource, is more particularly based on as micro- suitable for adjusting and stimulating the metabolic of application on human skin and hair follicle The extract of the plants such as algae.Especially, the purpose of the present invention is exploitation is respectively used to the new extraction of cosmetics and dept. of dermatology's application Thing, it is adjusted at the same time, is ie, meaning increased, improves and/or is stimulated

Melanin generation in ■ people's hair and skin；

■ grows or, alternatively, suppresses the growth of people's hair and hair follicle；

Collagen synthesis in the skin corium of ■ people；

Hyaluronic acid synthesis in the skin corium of ■ people；

The Keratinocyte differentiation of cuticula in the epidermis of ■ people and adjusting；

The stimulation of ■ cell proliferations, the more particularly stimulation to melanocyte proliferation；

The improvement of ■ wound healings, the more particularly stimulation to fibroblast and keratinocyte proliferation；And

The improvement of ■ lipolysis.

The purpose of the present invention is the extract of microalgae, microalgae is selected from

(i) garlic Trentepohlia

(ii) Thalassiosira

(iii) Chaetoceros category, and/or

(iv) Chlorococcum

The extract can be by using selected from C₁-C₄Aliphatic alcohol, ethyl acetate, the solvent processing of water or their mixture are described micro- Algae, the extract of dissolving is removed from residue and is obtained from the pure extract of the solvent recovery.

, it is surprising that in terms of the adjusting of the skin and hair follicle of desired people, especially with respect to melanogenesis, hair The growth of capsule, or alternatively, hair growth inhibition, the synthesis of collagen and hyaluronic acid, Keratinocyte differentiation, melanocyte In terms of cell Proliferation and effect related to this, the improvement of wound healing and the improvement of last lipolysis, when with from When the product that in the market obtains is compared, it was observed that said extracted thing shows excellent performance.The present invention covers the property of extract Speech that can be closely related from the property of different extractants.In other words, different solvents causes extract to have different groups Compound and different property.

Microalgae

According to the present invention, the microalgae for having identified four types is suitable for solving above-mentioned complicated brief description.

Garlic Trentepohlia

Garlic Trentepohlia (Monodus) belongs to true eyespot algae guiding principle (Eustigmatophyceae), it is represented containing abundant more insatiable hungers With the microalgae of aliphatic acid.Preferable strain is：Garlic algae (also referred to as subterranea Monodopsis), preferably using especially From the algae and original managed by Scotland Marine Sciences association (Scottish Association for Marine Science) The strain CCAP 848/1 that lively thing culture collection obtains (is also registered as ATCC 30593 in other collections；UTEX 151 and SAG 848-1).

Thalassiosira

In the various strains of the hailian seaweed of Bacillariophyceae are also belonged to, Thalassiosira pseudonana (Thalassiosira pseudonana) is It is most commonly known in the centriate diatom of ocean.It is chosen as the first eucaryon phytoplankton of genome sequencing.Thalassiosira pseudonana This research is selected for, is widely distributed in because it is the model of diatom physiological Study and belonging in the ocean of All Around The World Category, and have relatively small genome in 34,000,000 base-pairs.Especially, from Australian Union's science and industrial research The strain CS173 that tissue preservation center obtains (is also registered as at Provasoli-Guillard national marine cultures center CCMP1335, is N EPCC58 at Canadian culture of microorganism center) it is preferred for our purpose.The clone clon is initial Collected, and continuously preserved in the medium from this gulf (Long Island, New York) of Morici in 1958.

Wei Shi hailian seaweeds (TW) are a kind of big diatoms (8-15 μm of 6-20 μ ms) used in shrimp and shellfishery.It is this Algae is considered the best algae for juvenile prawn by some farms.Its cell size is 16 times and four of Chaetoceros biomass 3 times of slit bamboo or chopped wood algae biomass.This algae is measured as about 15 microns but is contracted to about 5 microns in summer in the winter time.According to culture The amount of Determination of Chlorophyll, the color of TW change from brown to green to yellow.The change of this color not shadow in any way Ring the quality of algae.The strain of all sea chain microalgaes is adapted as starting material, to obtain extract according to the present invention.

Chaetoceros category

Chaetoceros, belong to Bacillariophyceae, it may be possible to which the maximum category of halomereid, more particularly diatom have the pact of description 400 kinds.Although these numerous descriptions are all no longer sound.Under normal conditions, be very difficult between different Chaetoceros algaes with Distinguish.Have been carried out repeatedly attempting this big category reassembling into subgenus, and this work is still underway.However, due to this Most of in a little work for making great efforts description species concentrate on northern area, and it is global to belong to, it is possible that having substantial amounts of Tropical species are not still described.Compilation illustrates the relevant suitable Chaetoceros strain of the present invention below：

■ exceptions Chaetoceros (Chaetoceros abnormis A.I.Proshkina-Lavrenko)

■ thorn Chaetoceros (Chaetoceros aculeatus I.V.Makarova)

■ A Deli Chaetoceros (Chaetoceros adelianus E.E.Manguin)

■ gradually bent Chaetoceros (Chaetoceros aduncus I.N.Sukhanova)

■ equalization Chaetoceros South Pole mutation (Chaetoceros aequatorialis var.antarcticus Manguin)

■ equalizations Chaetoceros (Chaetoceros aequatorialis Cleve)

The narrow gap Chaetoceros of ■ intend symmetrical variant (Chaetoceros affinis f.pseudosymmetricus (E.Steemann Nielsen)M.Torrington-Smith)

The parallel modification of the narrow gap Chaetoceros of ■ (Chaetoceros affinis f.parallelus M.Thorrington- Smith)

The narrow gap Chaetoceros of ■ do not wait modification (Chaetoceros affinis f.inaequalis M.Thorrington- Smith)

The narrow gap Chaetoceros of ■ (Chaetoceros affinis Lauder)

■ goose cream Chaetoceros (Chaetoceros amanita A.Cleve-Euler)

■ bridgings Chaetoceros (Chaetoceros anastomosans Grunow)

■ has rib Chaetoceros (Chaetoceros angularis Schutt)

■ has angle Chaetoceros (Chaetoceros angulatus F.Schutt)

■ bridging Chaetoceros beauties mutation (Chaetoceros anostomosans var.speciosus F.Schutt)

■ is by first Chaetoceros (Chaetoceros armatus T.West)

■Chaetoceros astrabadicus A.Henckel

■ Atlantic Ocean Chaetoceros tight type mutation (Chaetoceros atlanticus var.compactus (F.Schutt) P.J.Cleve)

■ Atlantic Ocean Chaetoceros Naples mutation (Chaetoceros atlanticus var.neapolitanus (Schroeder)Hustedt)

■ Atlantic Ocean Chaetoceros enlargement type mutation (Chaetoceros atlanticus var.tumescens A.Grunow)

■ Atlantic Ocean Chaetoceros (Chaetoceros atlanticus Cleve)

The bold Chaetoceros modification in the ■ Atlantic Ocean (Chaetoceros atlanticus f.audax (F.Schutt) H.H.Gran)

■ Atlantic Ocean Chaetoceros cross mutation (Chaetoceros atlanticus var.cruciatus (G.Karsten) M.Thorrington-Smith)

The bold Chaetoceros of ■ (Chaetoceros audax F.Schutt)

■Chaetoceros bacteriastrius G.C Wallich

■ knurls face Chaetoceros imbricate modification (Chaetoceros bacteriastroides f.imbricatus (L.A.Mangin)M.Thorrington-Smith)

■ knurls face Chaetoceros (Chaetoceros bacteriastroides G.H.H.Karsten)

■Chaetoceros bermejense D.U.Herndndez-Becerril

■Chaetoceros bisetaceus J.Schumann

■ north Chaetoceros (Chaetoceros borealis J.W.Bailey)

■Chaetoceros borealoides H.L.Honigmann

The short Chaetoceros of ■ (Chaetoceros breve F.Schutt)

The short spore Chaetoceros of ■ (Chaetoceros brevis Schutt)

■Chaetoceros brussilowi A.Henckel

■ ox horn shape ChaetocerosG.H.H.Karsten(Chaetoceros buceros G.H.H.Karsten)

■ ox horn shape ChaetocerosKarsten(Chaetoceros buceros Karsten)

The spherical Chaetoceros of ■ (Chaetoceros bulbosus (Ehrenberg) Heiden)

The spherical Chaetoceros cross modifications of ■ (Chaetoceros bulbosus f.cruciatus (G.Karsten) H.Heiden)

■Chaetoceros bulbosus f.schimperana(G.Karsten)H.Heiden

■Chaetoceros bungei Honigmann

The short modification of the calcareous Chaetoceros of ■ (Chaetoceros calcitrans f.pumilus Takano)

■ California Chaetoceros (Chaetoceros californicus A.Grunow)

The thin lid Chaetoceros of ■ (Chaetoceros capense G.H.H.Karsten)

■ the Caspian Sea Chaetoceros (Chaetoceros caspicus C.E.H.Ostenfeld)

■ the Caspian Sea Chaetoceros, which add, draws mutation (Chaetoceros caspicus var.karianus A.Henckel)

■Chaetoceros caspicus f.pinguichaetus A.Henckel & P.Henckel

■ Cattell ChaetocerosKarsten(Chaetoceros castracanei Karsten)

■ Cattell ChaetocerosG.H.H.Karsten(Chaetoceros castracanei G.H.H.Karsten)

■Chaetoceros ceratospermus var.minor A.F.Meunier

■Chaetoceros ceratosporus var.brachysetus Rines & Hargraves

■ angles spore Chaetoceros (Chaetoceros ceratosporus Ostenfeld)

■Chaetoceros chunii G.H.H.Karsten

■ is around spore Chaetoceros (Chaetoceros cinctus Gran)

The bar-shaped Chaetoceros of ■ (Chaetoceros clavigera C.E.H.Ostenfeld)

The bar-shaped Chaetoceros of ■ (Chaetoceros clavigerus A.Grunow)

■ kirschner Chaetoceros (Chaetoceros clevei F.Schutt)

The close poly- Chaetoceros (Chaetoceros coarctatus Lauder) of ■

■Chaetoceros cochleus F.Schutt

The close Chaetoceros of ■ (Chaetoceros compactus F.Schutt)

The flat very thin mutation of face Chaetoceros of ■ (Chaetoceros compressus var.gracilis F.Hustedt)

■Chaetoceros compressus var.hirtisetus J.E.B.Rines & P.E.Hargraves

■Chaetoceros concavicorne Mangin

■ filiforms Chaetoceros (Choetoceros confervoides J.Rolfs)

■ mixes Chaetoceros (Choetoceros confusus S.L.VonLondinghom)

■ hangs contracting Chaetoceros Choetoceros constrictus Gran)

■ turns round Chaetoceros (Chaetoceros convolutus Castrocane)

■Chaetoceros convolutus f.trisetosus Brunei

■ turns round Chaetoceros dancing posture modification (Chaetoceros convolutus f.volans L.I.Smirnovo)

■ is preced with spore ChaetocerosG.Leuduger-Fortmorel(Choetoceros cornutus G.Leuduger- Fortmorel)

■ is preced with spore ChaetocerosGran(Chaetoceros coronotus Gran)

The double ridge Chaetoceros (Chaetoceros costatus Pavillord) of ■

■ knuckle-tooth Chaetoceros (Chaetoceros crenatus (C.G.Ehrenberg) T.Brightwell)

■ must shape Chaetoceros (Chaetoceros crinitus Schutt)

■Choetoceros criophilus Castrocane

The cross Chaetoceros Choetoceros cruciatus G.H.H.Karsten of ■)

■Chaetoceros curvotus Castrocane

■ rotation chain Chaetoceros (Chaetoceros curvisetus Cleve)

■ reaches base of a fruit Chaetoceros (Chaetoceros dadayi Pavillard)

■ Denmark Chaetoceros (Chaetoceros danicus Cleve)

The weak Chaetoceros of ■ (Chaetoceros debilis Cleve)

■ and base Chaetoceros unit cell modification (Chaetoceros decipiens f.singuloris H.H.Gran)

■ and base Chaetoceros (Chaetoceros decipiens Cleve)

The thin and delicate Chaetoceros of ■ (Chaetoceros delicatulus C.E.H.Ostenfeld)

The close even Chaetoceros (Chaetoceros densus Cleve) of ■

■ imperial crowns Chaetoceros (Choetoceros diadema (Ehrenberg) Gran)

■Choetoceros dichaeta f.unicellularis H.Heiden

The double fork Chaetoceros (Choetoceros dichaetus Ehrenberg) of ■

■Chaetoceros dichaetus var.polygonus(F.Schutt)H.Heiden

■Chaetoceros didymus var.praelongus E.J.Lemmermonn

■Chaetoceros didymus f.aestivus H.H.Gran

The double prominent Chaetoceros autumn modifications (Chaetoceros didymus f.autumnalis H.H.Gran) of ■

The double prominent Chaetoceros (Choetoceros didymus C.G.Ehrenberg) of ■

The intractable Chaetoceros of ■ (Chaetoceros difficilis Cleve)

Bis- row Chaetoceros of ■ (Choetoceros distichus F.Schutt)

■ distinctnesses Chaetoceros (Chaetoceros distinguendus E.J.Lemmermonn)

■Chaetoceros diversicurvatus Van Goor

The different angle Chaetoceros Mediterranean mutation of ■ (Chaetoceros diversus var.mediterraneus J.LB.Schroder)

The different angle Chaetoceros of ■ (Chaetoceros diversus Cleve)

■ Emhorns Chaetoceros (Chaetoceros eibenii (Grunow) Meunier)

■Chaetoceros elmorei Boyer

The elongated Chaetoceros of ■ (Chaetoceros elongatus Honigmann)

■Chaetoceros exospermus Meunier

The outer Chaetoceros (Chaetoceros externus Gran) of ■

■ intends Chaetoceros (Chaetoceros fallax Prosckina-Lavrenko)

The thick stock Chaetoceros of ■ (Chaetoceros femur F.Schutt)

■Chaetoceros filiferus G.H.H.Karsten

■ filiforms Chaetoceros (Chaetoceros filiforme Meunier)

■ bends Chaetoceros (Chaetoceros flexuosus Mangin)

■ fragments Chaetoceros (Chaetoceros fragilis Meunier)

The ■ fork long angle mutation of Chaetoceros (Chaetoceros furca var.macroceras J.LB.Schroder)

■ prongs Chaetoceros (Chaetoceros furcellatus J.W.Bailey)

■ shuttles Chaetoceros (Chaetoceros fusus F.Schutt)

■Chaetoceros galvestonense Collier & Murphy

■Chaetoceros gastridius(C.G.Ehrenberg)T.Brightwell

■Chaetoceros gaussii Heiden & Kolbe

■ glacial epoches Chaetoceros (Chaetoceros glacialis A.Henckel)

■ grignard Chaetoceros (Chaetoceros glandazii Mangin)

The smooth Chaetoceros of ■ (Chaetoceros gobii A.Henckel)

■ Chaetoceros gracilis (Chaetoceros gracilis Pantocsek)

■ lattice Shandong Chaetoceros (Chaetoceros grunowii F.Schutt)

■Chaetoceros hendeyi Manguin

■Chaetoceros hispidus var.monicae A.Grunow

■Chaetoceros hohnii Graebn.& Wujek

■ is without ditch Chaetoceros (Chaetoceros holsaticus Schutt)

■ heterodoxies Chaetoceros (Chaetoceros ikari B.V.Skvortzov)

■ spine Chaetoceros (Chaetoceros imbricatus Mangin)

The foldable Chaetoceros convex mutation of ■ (Chaetoceros incurvus var.umbonatus Castracane)

The foldable Chaetoceros of ■ (Chaetoceros incurvus Bailey)

■ India Chaetoceros Choetoceros indicus Karsten)

■Chaetoceros ingolfianus Ostenfeld

Chaetoceros (Chaetoceros intermedius A.Henckel) among ■

■, which adds, draws Chaetoceros (Chaetoceros karianus Grunow)

■Chaetoceros karyanus A.Henckel

■ grams of Buddhist nun's Chaetoceros (Chaetoceros knipowitschii A.Henckel)

■ hangs down edge Chaetoceros (Chaetoceros laciniosus Schut)

■ hangs down edge Chaetoceros protuberance modification (Chaetoceros laciniosus f.protuberans M.Thorrington- Smith)

■ hangs down edge Chaetoceros ocean modification (Chaetoceros laciniosus f.pelagicus H.H.Gran)

■ labor morals Chaetoceros (Chaetoceros lauderi Ralfs)

■Chaetoceros leve F.Schutt

■ streamsides beach Chaetoceros (Chaetoceros littorale litorale E.J.Lemmermann)

■ Rockwells Chaetoceros pincers mutation (Chaetoceros lorenzianus var.forceps A.F.Meunier)

■ Rockwells Chaetoceros (Chaetoceros lorenzianus Grunow)

■Chaetoceros malygini A.Henckel

Chaetoceros (Chaetoceros medius F.Schutt C) in the middle part of ■

■ south Chaetoceros (Chaetoceros meridiana (F.Schutt) G.Karsten)

■ plum Dun Shi Chaetoceros (Chaetoceros mertensii H.L.Honigmann)

The short fork Chaetoceros (Chaetoceros messanense Castracane C) of ■

The minimum Chaetoceros of ■ (Chaetoceros minimus (Levander) D.Marino, G.Giuffre, M.Montresor & A.Zingone)

■Chaetoceros misumensis H.H.Gran & K.Yendo

■ high spores Chaetoceros (Chaetoceros mitra (J.W.Bailey) Cleve)

■ tries to gain the double mutation of Le Shi Chaetoceros (Chaetoceros muelleri var.duplex E.J.Lemmermann)

■ tries to gain the wide salt mutation of Le Shi Chaetoceros (Chaetoceros muelleri var.subsalsum J.R.Johansen S.Rushforth)

■ Chaetoceros muelleris (Chaetoceros muelleri E.J.Lemmermann)

■ tries to gain Le Shi Chaetoceros salt pool mutation (Chaetoceros muellerii var.subsalsus J.R.Johansen Rushforth)

■Chaetoceros nansenii A.Henckel

■ swims Chaetoceros (Chaetoceros natatus E.E.Manguin)

■ piebalds Chaetoceros (Chaetoceros neglectus Karsten)

■Chaetoceros neobulbosus T.V.Desikachary,S.Gowthaman & Y.Latha

■Chaetoceros neocompactus S.L.VanLandingham

The new elongated Chaetoceros (Chaetoceros neogracile S.L.VanLandingham) of ■

■Chaetoceros neupokojewii A.Henckel

■ Japan Chaetoceros Choetoceros nipponicus J.Ikori)

■ tool tooth Chaetoceros (Chaetoceros odontella (CG.Ehrenberg) G.L.Rabenhorst)

■ hilllocks village Chaetoceros tetrasetus mutation (Chaetoceros okamurae var.tetrasetus J.Ikari)

■ hilllocks village Chaetoceros (Chaetoceros okamurae J.Ikari)

■Chaetoceros ostenfeldii P.T.Cleve

■Chaetoceros pachtussowii A.Henckel

■Chaetoceros pachyceros R.Margalef

■ Pacific Ocean Chaetoceros (Chaetoceros pacificus J.Ikari)

The unusual Chaetoceros of ■ (Chaetoceros paradoxus Cleve)

■Chaetoceros paradoxus var.luedersii Engler

The small Chaetoceros of ■ (Chaetoceros parvus F.Schutt)

■ ditches life Chaetoceros runic modification (Chaetoceros paulsenii f.robustus A.Henckel)

■Chaetoceros pavillardii J.Ikari

■ oceans Chaetoceros (Chaetoceros pelagicus)

■ overhangs Chaetoceros (Chaetoceros pendulus Karsten)

The small Chaetoceros of ■ (Chaetoceros perpusillus Cleve)

■ Peru Chaetoceros tie up more mutation (Chaetoceros peruvianus var.victoriae Karsten)

The very thin mutation of ■ Peru Chaetoceros (Chaetoceros peruvianus var.gracilis J.LB.Schroder)

■ Peru Chaetoceros (Chaetoceros peruvianus Brightwell)

The thick mutation of ■ Peru Chaetoceros (Chaetoceros peruvianus var.robustum P.T.Cleve)

■Chaetoceros peruvianus var.suadivae Karsten

■ Peru Chaetoceros dancing posture modification (Chaetoceros peruvionus f.volans (F.Schutt) C.E.H.Ostenfeld)

■ Peru Chaetoceros runic modification (Chaetoceros peruvianus f.robustus (P.T.Cleve) C.E.H.Ostenfeld)

General lucky Chaetoceros (Chaetoceros phuketensis J.E.B.Rines, the P.Boonruang ＆ E.C of ■ Theriot)

The stout and strong Chaetoceros of ■ (Chaetoceros pingue A.Henckel)

■Chaetoceros pinguichaetus A.Henckel & P.Henckel

The new generation Chaetoceros of ■ (Chaetoceros pliocenus J.-J.Brun)

■ protuberance Chaetoceros (Chaetoceros protuberans H.S.Lauder)

■ intends Ovshinsky Chaetoceros (Chaetoceros pseudoaurivillii J.Ikari)

■ sends out shape Chaetoceros (Chaetoceros pseudocrinitus Ostenfeld)

■ intends rotation chain hair algae (Chaetoceros pseudocurvisetus Mangin)

The false double thorn Chaetoceros (Chaetoceros pseudodichaeta J.Ikari) of ■

■Chaetoceros pundulus G.H.H.Karsten

■ radiation Chaetoceros (Chaetoceros radians F.Schutt)

■ root shapes Chaetoceros (Chaetoceros radicans F.Schutt)

■ long sheath Chaetoceros runics modification (Chaetoceros recurvatus f.robustus Henckel)

■ long sheaths Chaetoceros (Chaetoceros recurvatus Henckel)

■ runics Chaetoceros (Chaetoceros robustus (P.J.Cleve) C.E.H.Ostenfeld)

■ mouth shapes Chaetoceros (Chaetoceros rostratus Lauder)

■Chaetoceros russanowi A.Henckel

■ happiness salt Chaetoceros (Chaetoceros salsugineus Takano)

■ ring-types Chaetoceros (Chaetoceros saltans P.J.Cleve)

■ Heidi Schmids Chaetoceros (Chaetoceros schmidtii C.E.H.Ostenfeld)

The black Chaetoceros strand modifications of ■ (Chaetoceros schuettii f.oceanicus H.H.Gran)

■ Chaetoceros (Chaetoceros secundus P.T.Cleve)

■ chains thorn Chaetoceros (Chaetoceros seiracanthus Gran)

■ stockless Chaetoceros (Chaetoceros sessile Gr0ntved)

■Chaetoceros setoense J.Ikari

■ plug tongue ear Chaetoceros (Chaetoceros seychellarus G.H.H.Karsten)

■ plug tongue ear Chaetoceros Australia modification (Chaetoceros seychellarus var.austral E.E.Manguin)

■ Siam Chaetoceros (Chaetoceros siamense C.E.H.Ostenfeld)

The similar Chaetoceros of ■ (Chaetoceros similis Cleve)

The simple Chaetoceros of ■ (Chaetoceros simplex C.E.H.Ostenfeld C)

■ bone bars Chaetoceros (Chaetoceros skeleton F.Schutt)

■ consor Chaetoceros radiation modification (Chaetoceros socialis f.radians (F.Schutt) A.I.Proshkina-Lavrenko)

■ consor Chaetoceros (Chaetoceros socialis Lauder)

■ consor Chaetoceros autumn mutation (Chaetoceros socialis var.autumnalis Prosckina- Lavrenko)

■Chaetoceros sedowii A.Henckel

The straight Chaetoceros of ■ (Chaetoceros strictus G.H.H.Karsten)

The sub- flat Chaetoceros (Chaetoceros subcompressus J.L.B.Schroder) of ■

■ salt pool Chaetoceros (Chaetoceros subsalsus Lemmermann)

■ crown spores Chaetoceros (Chaetoceros subsecundus (Grunow ex Van Heurck) Hustedt)

The thin and delicate Chaetoceros of ■ (Chaetoceros subtilis Cleve)

■ Sumateras Chaetoceros (Chaetoceros sumatranus Karsten)

The very thin Chaetoceros of ■ (Chaetoceros tenuissimus A.F.Meunier)

■ cylinder Chaetoceros spinules modification (Chaetoceros teres f.spinulosus H.H.Gran)

■ cylinders Chaetoceros (Choetoceros teres Cleve)

■Choetoceros tetrachaeta Ehrenberg

Tetra- Chaetoceros of ■ (Chaetoceros tetras G.H.H.Karsten)

Tetra- stupefied Chaetoceros of ■ (Choetoceros tetrastichon Cleve)

■ Shan Shi Chaetoceros (Chaetoceros thienemannii Hustedt)

■Chaetoceros throndsenii var.trisetosus Zingone

■Chaetoceros throndsenii var.throndsenia D.Marino,M.Montresor & A.Zingone

■Chaetoceros throndsenii(Marino,Montresor,& Zingone)Marino,Montresor & Zingone

■ turns round chain Chaetoceros (Chaetoceros tortissimus H.H.Gran)

■Chaetoceros transisetus J.R.Johansen & J.S.Boyer

■ Fan Shi Chaetoceros (Chaetoceros vanheurckii H.H.Gran)

■ worm limbs Chaetoceros (Chaetoceros vermiculus F.Schutt)

■ drapes over one's shoulders a mao Chaetoceros (Chaetoceros villosus Kutzing)

■ Wei Sidula Chaetoceros (Chaetoceros vistulae C.Apstein)

■ dancing postures Chaetoceros (Chaetoceros volans F.Schutt)

■ Wei Shi Chaetoceros (Chaetoceros weissflogii F.Schutt)

■Chaetoceros wighamii Brightwell

■ Websters Chaetoceros (Chaetoceros willei Grunow)

■ pricks card Chaetoceros lunge mutation (Chaetoceros zachariasi var.longus H.L.Honigmann)

■ pricks the variegated mutation of card Chaetoceros (Chaetoceros zachariasii var.variatus H.L.Honigmann)

■ pricks card Chaetoceros avris mutation (Chaetoceros zachariasii var.latus H.L.Honigmann)

■ pricks card Chaetoceros (Chaetoceros zachariasii Honigmann)

■Chaetoceros ziwolkii A.Henckel

Two kinds of different strains are have studied for laboratory report herein：The first is the unknown Chaetoceros category in source, and second The short modification of calcareous Chaetoceros (Chaetoceros calcitrans f.pumilus), its be in nineteen sixty from Separated ocean algae strain in the seawater of Pu peace (Japan, Chiba area) near Umbayashi.The latter is by Scotland Marine Sciences Association manages algae and protozoan culture collection (CCAP) is achieved and (also registered for CCAP 1010/11 in other collections For PLY537；CCMP1315；NEPCC 590).

Chlorococcum

Chlorococcum is the category in Chlorococcum section.Compilation illustrates the relevant suitable Chlorococcum strain of the present invention below：

■ acid Chlorococcum (C.acidum)

■ dragon's paws Chlorococcum (C.aegyptiacum)

■ botryoidalis Chlorococcum (C.botryoides)

■C.choloepodis

■ lemon shape Chlorococcums (C.citriforme)

■C.costatozygotum

■ binarys Chlorococcum (C.diplobionticum)

■ drastic cracks Chlorococcum (C.dissectum)

■C.echinozygotum

■C.elbense

■C.elkhartiense

■ ellipses Chlorococcum (C.ellipsoideum)

■ chroococcoids (C.hypnosporum)

■ dipping Chlorococcums (C.infusionum)

■C.isabeliense

■ is shallow to split Chlorococcum (C.lobatum)

■C.mocrostigmatum

The minimum Chlorococcums of ■ (C.minimum)

The small Chlorococcums of ■ (C.minutum)

■C.novae-angliae

■C.oleofaciens

■ olives Chlorococcum (C.olivaceum)

■C.pamirum

■C.pinguideum

■ multiforms Chlorococcum (C.polymorphum)

■ puppet glue net Chlorococcums (C.pseudodictyosphaerium)

■C.pyrenoidosum

The soft Chlorococcums of ■ (C.refringens)

■ salt ponds Chlorococcum (C.salinum)

■ splits wall Chlorococcum (C.schizochlamys)

■C.schworzii

The sub- extra large Chlorococcums (C.submarinum) of ■

■ hemps Chlorococcum (C.tatrense)

■C.vacuolatum

For this research, respectively using two kinds of different algae strains：The first is the unknown Chlorococcum in source, and is for second micro- Small Chlorococcum, a kind of freshwater microalgae, algae and protozoan culture collection by the management of Scotland Marine Sciences association (CCAP) achieve as algae strain CCAP213/7 (SAG213-7；SAG21.95；TACC117；UTEX290).The latter is by Bold from India Soil isolate.

Small Chlorococcum be currently viewed as following species classification synonym (Guiry, M.D.＆ Guiry, G.M.2011.AlgaeBase.World-wide electronic publication,National University of Ireland,Galway.http://www.algaebase.org；searched on 07 Septem ber 2011)：

Coarse Chlorococcum (Chlorococcum scabellum Deason＆Bold 1960)

Golden Chlorococcum (Chlorococcum aureum Archibald＆Bold 1970)

Chlorococcum reticu latum Archibald& Bold 1970

Chlorococcum sphacosum Archibald& Bold 1970

Chlorococcum typicum Archibald& Bold 1970

Extraction process

Another object of the present invention is related to acquisition

(i) garlic Trentepohlia,

(ii) Thalassiosira,

(iii) Chaetoceros category, and/or

(iv) Chlorococcum

Extract method, this method comprises the following steps：

(a) optionally at elevated temperatures, the microalgae is made with suitably causing being selected from for the active amount for being moved to solvent phase C₁-C₄The solvent contact of aliphatic alcohol, ethyl acetate, water or its mixture,

(b) extract of dissolving is removed from residue, and

(c) from the pure extract of the solvent recovery.

Substantially, extract according to the present invention can be produced by method known per se, for example, by using foregoing description Solvent carry out aqueous solution, organic solution or water/the organic solvent extraction of microalgae.Suitable extraction process carries to be any traditional Taking technique：As soaked, soaking again, digesting, stirring immersion, extraction of ocean eddies, ultrasonic extraction, adverse current are extracted, are percolated, are percolated again, taken out (the lower extraction of decompression) is proposed, solid/liquid extracts under diacolation and continuous backflow.It is favourable that diafiltration, which is used for industrial use,.It can make The method that any size known to expert reduces, for example, freeze grinding.The preferable solvent of extraction process be methanol, ethanol, Isopropanol, ethyl acetate and water (being preferably the hot water of more than 80 DEG C of temperature, particularly more than 95 DEG C) or the organic solvent With the mixture of water, more particularly, there is the alcohol of the low molecular weight of high or low water content.Particularly preferably with methanol, The extraction of ethanol and their aqueous mixture.Extraction process generally temperature be from about 20 to about 100 DEG C, be preferably from about 50 to Carried out at about 70 DEG C.In a preferred embodiment, extraction process carries out in inert gas atmosphere, to avoid extracting solution In component oxidation.This temperature more than 40 DEG C is even more important in the case of being extracted.By expert according to starting material, The selective extraction time such as ratio of extraction process, Extracting temperature, solvent and raw material.After extraction process, the coarse extraction of acquisition Thing is alternatively subjected to other exemplary steps, such as, for example, purifying, concentration and/or discoloration.If desired, preparation carries Thing is taken to be subjected to, for example, optionally removing individually unwanted component.Although can proceed to any degree in extraction process, But usually it is continuing to and exhausts.Typical yield (number of the dry matter based on the raw material used of=extraction in the extraction of starting material Amount) it is from about 1 to about 50%, preferably from about 2 to about 20%, more preferably from about 5 to about 10% (weight)-calculated with starting material Amount.

Hereinafter, three kinds of typical process for obtaining extract according to the present invention are described in further detail：

Methanol or ethanol or isopropanol or ethyl acetate extraction process：Single solvent extraction

Handled at room temperature with 100 milliliters of solvent in the dark, carry out extracting every gram of dry biomass when stirred suspension 16 is small；

By being centrifuged 15 minutes in 2000G, remaining cell material is separated from extract；

By suspending in 50 milliliters of solvents, the biomass of wash residual；

By being centrifuged 15 minutes in 2000G, cellular material is separated from cleaning solvent；

Suspend again by 50 milliliters of solvents, the biomass of wash residual；

By being centrifuged 15 minutes in 2000G, cell material is separated from cleaning solvent；

The extract and the amount of cleaning solvent that mixing first is collected, it is micro- that the extracting solution of gained is considered to have conventional concentration 5000 Grams per milliliter (μ g/ml) (the drying algaes of 1000 micrograms in 200 milliliters of solvents).

Two steps are continuously extracted：Ethanol>Water

In order to obtain the separation of the first time between more lipophilic compound and hydrophilic compound, pass through such as single solvent extraction It is described that dry Algal material is handled with ethanol, then handle remaining cell material again by the use of water as solvent Material, implements the extraction experiment of two steps.The experimental program of description provides the identical cell material with different solvent continuous processings Sample, but the first extract in only described continuous processing is obtained from complete cell material；Just because of this, it is referred to as " directly extracting " (in this case, direct ethanol extraction).The subsequent extracted thing prepared from remaining cell material, progressively obtains Less compound, hereinafter referred to as " continuous (sequential) extract " (in this case, continuous water extraction Thing).

Three steps are continuously extracted：Ethyl acetate>Absolute ethyl alcohol>Water

Implement the extraction experiment of three steps, to obtain preferable separation of the lipophilic compound from weak lipophilicity and hydrophilic compound. Technical operation is identical with the extraction experiment for being used for single solvent extraction of description, but in this case, with ethyl acetate execution the Once extract, then extraction process is repeated twice completely, each process change Extraction solvent (ethyl acetate>Ethanol>Water).

Microalgae component contains very abundant reactive compound, is adapted to produce a contrary effect on same destination organization sometimes. As a result, the bioactivity of every kind of extract is strongly depend on the strain of microalgae algae and the extraction process used., it is surprising that For example, produce using direct ethanol or direct method of isopropanol extraction Chaetoceros cell material and suppress unnecessary hair growth Extract, and the continuous ethanol extract obtained according to three step continuous extractions from identical cell material shows opposite effect Fruit, i.e. the extract hair follicle stimulating grow.

Commercial Application

Another object of the present invention is to be directed to cosmetic composition, it includes the extract of microalgae and cosmetically acceptable Auxiliary material, the microalgae are selected from

(i) garlic Trentepohlia,

(ii) Thalassiosira,

(iii) Chaetoceros category, and/or

(iv) Chlorococcum,

The cosmetically acceptable auxiliary material is selected from C₁-C₄Aliphatic alcohol, polyalcohol, oil ingredient, the water with 3 to 12 carbon atoms With their mixture.Suitable auxiliary material includes, for example, ethanol, propyl alcohol, isopropanol, the butanol of all isomeric forms, second two Alcohol and/or propane diols and its dimer and tripolymer, glycerine, glucose, pentaerythrite etc..Disclosed in following chapters and sections Suitable oil component.

Composition can be included from 0.001 to 35, preferably from 0.5 to 20, more preferably from 2 to 10% weight-according in extract The amount that dry matter calculates.Remaining part is auxiliary material.Under normal conditions, extract local application；It is also possible, however, to use extraction Thing-after particularly encapsulating-is used for oral absorption.

Cosmetic composition

Cosmetic composition also includes dermatological compositions, and is particularly useful for the treatment of the skin of people and the composition of hair. The composition may include other compound, such as, for example, surfactant, oil body, emulsifying agent, superfatting agent, pearlescent waxes, Consistency factor, polymer, organo-silicon compound, wax, stabilizer, dandruff removing agent, biological agent, film forming agent, preservative, perfumery oil, Dyestuff etc. is as additional auxiliary agent and additive.

Surfactant

Other preferable auxiliary agents and additive are anion surfactant and/or amphoteric surfactant or amphion surface Activating agent.The typical example of anion surfactant be soap, alkylbenzenesulfonate, alkylsulfonate, alkene sulfonate, Alkylether sulfonate, glycerol ether sulfonate, methyl ester sulfonates, sulfo-fatty acid, alkyl sulfate, fatty alcohol ether sulphate, Glycerol ether sulfate, fatty acid ether sulfate, hydroxyl mixing ether sulfate, monoglyceride (ether) sulfate, fatty acid amide (ether) sulfate, list-and dialkyl sulfosuccinates, list-and dialkylsulfosuccinic acid amides hydrochlorate, three acid glycerol of sulfo group Ester, acid amides soap, ether carboxylic acid and its salt, fatty acid isethionates, fatty acid sarcosinates, fatty acid taurides, N- acyls Base amino acid：Such as, for example, acyl-lactate, acyl group tartaric acid salt, acyl glutamate, acylaspartic acid salt, alkyl are low Polyglucoside sulfate, protein fatty acid condensation product (victual for being based particularly on wheat) and alkyl (ether) phosphate. If anion surfactant contains polyglycol ether chain, they can have traditional homologue distribution, but their preferred tools The homologue for having narrow range is distributed.The typical example of both sexes or zwitterionic surfactant is alkyl betaine, alkyl Amido betaine, aminopropionate, amino glycine salt, imidazolinium betaine and sulfobetaines.All tables being previously mentioned Face activating agent is known compound.Their structure and the information of production can be found in relevant paper, referring to J.Falbe (chief editor), " Surfactants inConsumer Products ", Springer Verlag, Berlin, the 1987, the 54th Page is to page 124 or J.Falbe (chief editor), " Katalysatoren, Tenside und Mineraloladditive (catalysis Agent, surfactant and mineral oil additive) ", Di Mu publishing houses, Stuttgart, 1978, the 123-217 pages.In terms of preparation Calculate, degree of the surfactant in the preparation can be preferably 0.5 to 5% (weight from 0.1 to 10% (weight) Amount).

Oil ingredient (being also auxiliary material)

Forming the suitable oil body of cosmetically acceptable auxiliary material is, for example, being based on having 6 to 18 preferably 8 to 10 carbon atoms Aliphatic alcohol Guerbet alcohol, the C of straight chain₆-C₂₂The C of aliphatic acid and straight or branched₆-C₂₂The ester of fatty alcohol, or side chain C₆-C₁₃The C of carboxylic acid and straight or branched₆-C₂₂The ester of aliphatic acid：Such as, for example, myristyl myristate, palmitic acid Pork and beans Cool ester, stearic acid myristin, isostearic acid myristin, oleic acid myristin, behenic acids myristin, erucic acid nutmeg Ester, cetyl myristate, cetin, cetyl stearate, isostearic acid cetyl, cetyl ester, behenic acids Cetyl, erucic acid cetyl, myristic acid octadecane ester, palmitic acid octadecane ester, stearic stearolactone, isostearic acid 18 Alkyl ester, oleic acid octadecane ester, behenic acid octadecanes ester, erucic acid octadecane ester, iso stearyl myristinate, iso stearyl palm It is acid esters, iso stearyl stearate, iso stearyl isostearate, iso stearyl oleate, iso stearyl behenic acid ester, different hard Aliphatic radical oleate, myristic acid oleyl alcohol ester, palmitic acid oil alcohol ester, stearic acid oleyl alcohol ester, isostearic acid oleyl alcohol ester, oleic acid oleic alcohol It is ester, behenic acids oleyl alcohol ester, oleyl erucate, myristic acid behenyl alcohol ester, palmitic acid behenyl alcohol ester, Ying resin acid behenyl alcohols ester, different Hard resin acid behenyl alcohol ester, oleic acid behenyl alcohol ester, behenic acid behenyl alcohol ester, erucic acid behenyl alcohol ester, myristic acid savoy alcohol ester, palm fibre Palmitic acid acid savoy alcohol ester, stearic acid savoy alcohol ester, isostearic acid savoy alcohol ester, oleic acid savoy alcohol ester, behenic acid wooden dipper Dish alcohol ester, erucic acid savoy alcohol ester.Also suitably straight chain C₆-C₂₂- aliphatic acid and branched-chain alcoho (especially 2-Ethylhexyl Alcohol) Ester, C₁₈-C₃₈The C of-alkyl hydroxy carboxylic acid and straight or branched₆-C₂₂The ester of-fatty alcohol, especially apple dioctyl phthalate, straight chain And/or the aliphatic acid of side chain and polyalcohol (such as, for example, propane diols or two polyalcohols or three polyalcohols) and/or Guerbet alcohol Ester, based on C₆-C₁₀The triglyceride of-aliphatic acid, based on C₆-C₁₈- monoglyceride/diglyceride of the liquid of aliphatic acid/ Triglyceride mixture, C₆-C₂₂- fatty alcohol and/or Guerbet alcohol and the ester of aromatic carboxylic acid's (especially benzoic acid), C₂- C₁₂The alcohol with 1 to 22 carbon atom of-dicarboxylic acids and straight or branched or with 2 to 10 carbon atoms and 2 to 6 hydroxyl bases The ester of the polyalcohol of group, vegetable oil, primary alconol, hexamethylene, straight chain and the side chain C of substitution of side chain₆-C₂₂- fatty alcohol carbonates, it is all Such as, for example, carbonic acid dioctyl ester (CC), based on the fatty alcohol with 6 to 18 preferably 8 to 10 carbon atoms Guerbet carbonic esters, benzoic acid and straight chain and/or the C of side chain₆-C₂₂-ol (such asTN ester), straight or branched , symmetrically or non-symmetrically there is dialkyl ether per alkyl group 6 to 22 carbon atoms, such as, such as dioctyl ether (trade nameOE), the open-loop products of epoxidized fatty acid ester and polyalcohol, silicone oil (cyclomethicone, polysiloxanes methyl silicon Oily grade, etc.), aliphatic or alicyclic hydro carbons, such as, for example, saualane, squalene or dialkycyclohexane, and/or mineral Oil.

Emulsifying agent

Also other surfactants can be added in preparation as emulsifying agent, comprising for example：

The moles of ethylene oxide of ■ 2 to 30 and/or 0 to 5 mole propylene oxide and linear C_8-22Aliphatic alcohol, with C_12-22Aliphatic acid and With the addition product of the alkylphenol containing 8 to 15 carbon atoms in alkyl group；

The C of the mole ethylene oxides of ■ 1 to 30 and the addition compound product on glycerine_12/18Fatty-acid monoester and diester；

■ contains the monoglyceride of saturation and the undersaturated aliphatic acid and its ethyleneoxide addition product of 6 to 22 carbon atoms With diester and anhydrosorbitol monoester and diester；

The mole ethylene oxides of ■ 15 to 60 and castor oil and/or the addition compound product of rilanit special；

■ polyol esters, and especially, polyglycerol ester, such as, for example, polyglycerol polyricinoleate, the poly- 12- of polyglycereol Hydroxy stearic acid ester or polyglycereol dimeric dibasic acid isostearate.The mixture of some of such compound is also suitable；

The mole ethylene oxides of ■ 2 to 15 and castor oil and/or the addition compound product of rilanit special；

Cs of the ■ based on straight chain, side chain, insatiable hunger and/or saturation_6/22Aliphatic acid, castor oil acid and 12- hydroxy stearic acids and glycerine, Polyglycereol, pentaerythrite, dipentaerythritol, sugar alcohol (such as sorbierite), alkyl glucoside (such as methyl glucosamine, butyl Portugal Glucosides, lauryl glucoside) and polyglucoside (such as cellulose) partial ester；

■ monoalkyl phosphoric acid esters, dialkyl phosphate and trialkyl phosphates, and single-PEG- alkyl phosphates, two-PEG- alkyl Phosphate and/or three-PEG- alkyl phosphates, and their salt；

■ wool wax alcohols；

■ polysiloxanes/poly- alkyl, polyether copolymer and corresponding derivative；

■ pentaerythrites, aliphatic acid, the mixed ester of citric acid and fatty alcohol, and/or C_6-22Aliphatic acid, methyl glucoside and polynary The mixed ester of alcohol (preferably glycerine or polyglycereol),

■ polyalkylene glycol and

■ glycerol carbonates.

Ethylene oxide and/or propylene oxide and fatty alcohol, aliphatic acid, alkyl phenol, the monoglyceride and diester of aliphatic acid and dehydration Sorbitan monoesters and diester, or the addition compound product of castor oil is known commercial product.They are average degree of alkoxylation pair Should be in the ratio homologous mixture between the quantity of ethylene oxide and/or propylene oxide and the substrate of progress addition reaction.Epoxy The C of the addition compound product of ethane and glycerine_12/18Fatty-acid monoester and diester are the lipid layer enhancings for being known as cosmetic formulations Agent.

Typical anion emulsifier is aliphatic C_12-22Aliphatic acid, such as, for example, palmitic acid, stearic acid or behenic acid, and C_12-22Dicarboxylic acids, such as azelaic acid or decanedioic acid.Other suitable emulsifying agents are zwitterionic surfactants.Amphion table Face activating agent is that at least one quaternary ammonium group and the surface-active of at least one carboxyl and a sulfonate groups are included in molecule Compound.Specially suitable zwitterionic surfactant is the material of referred to as glycine betaine, as N- alkyl-N, N- Dimethyl Ammonium is sweet Propylhomoserin salt (N-alkyl-N, N-dimethyl ammonium glycinat), for example, cocodimethyl ammonium glycinate, N- cocamidopropyls-N, N- dimethylammonium glycinates, for example, cocoamidopropyl dimethyl ammonium glycinate, alkyl or 2- alkyl -3- carboxymethyls -3- hydroxyethyl imidazoles quinoline and cocoamidoethyl carboxymethyl hydroxyethyl containing 8 to 18 carbon atoms in acyl group Methylglycine salt.Particularly preferably spread out according to the fatty acid amide of the CTFA known Cocoamidopropyl betaines named Biology.Amphoteric surfactant is also suitable emulsifying agent.Amphoteric surfactant is the compound of surface-active, its except C_8/18Beyond alkyl or carboxyl groups, in molecule containing at least one free amine group and at least one-COOH- or- SO₃H- groups, and inner salt can be formed.The example of suitable amphoteric surfactant be N- alkyl glycines, N- alkylpropionic acids, N- alkyl aminos butyric acid, N- alkyliminodipropionics, N- ethoxy-N- alkylamide propyls glycine, N- alkyl taurines, P dialkylaminobenzoic acid containing about 8 to 18 carbon atoms in N- alkylsarcosines, 2- alkyl aminopropionic acids and alkyl.It is particularly preferred Amphoteric surfactant is N- coco alkyl amino propionic acids salt, cocoamidoethyl aminopropionate and C_12/18Acyl group flesh ammonia Acid.

Superfatting agent

Superfatting agent can be selected from these materials, for example, lanolin, lecithin and polyethoxylated and acylated lanolin and Lecithin derivative, polyol fatty acid ester, monoglyceride and Marlamid, also serve as the fat of foam stabiliser Fat acid alkanolamide.

Consistency factor

Main consistency factor (consistency factors) to be used is containing 12 to 22 and preferably 16 to 18 carbon atoms Aliphatic alcohol or hydroxy fatty alcohols and partial glyceride, aliphatic acid or hydroxy fatty acid.Preferably using these materials and alkyl Oligosaccharides glycosides and/or aliphatic acid N- methyl glucose amides and/or the poly- 12- hydroxy stearates of polyglycereol with identical chain length The combination of acid esters.

Thickener

Suitable thickener is the thickener of polymerization, such as：Type (hydrophilic silicon oxides), polysaccharide, particularly It is the phase of xanthans, guar gum, agar, alginates and methylcellulose, carboxymethyl cellulose and hydroxyethyl cellulose, aliphatic acid To high molecular weight polyethylene glycol monoesters and diester, polyacrylate (such as[Goodrich]or[Sigma]), polyacrylamide, polyvinyl alcohol and polyvinylpyrrolidone, surfactant：Such as, example Such as, ethoxylated fatty acid ester, aliphatic acid and polyalcohol (such as pentaerythrite or trimethylolpropane, close limit fat Alcohol ethoxylate) ester, and electrolyte：Such as sodium chloride and ammonium chloride.

Polymer

Suitable cationic polymer is, for example, cationic cellulose derivative, such as, for example, the title purchased from Amerchol For polymer JRQuaternized hydroxyethyl cellulose, cationic starch, diallyl ammonium salt and acrylamide are total to Polymers, quaternized vinyl pyrrolidone/vinylimidazole polymers, such as, for example,(BASF), poly- second The condensation product of two alkohol and amines, quaternized collagen polypeptide, such as, for example, Lauryldimonium Hydroxypropyl base hydrolyzes glue Former albumen (L, Grunau), quaternized Semen Tritici aestivi polypeptide, polyethyleneimine, cation type organic silicon polymer, Such as, for example, amino polydimethyl siloxane (amodimethicone), adipic acid and dimethylamino hydroxypropyl divinyl three Amine copolymer (Sandoz companies), the copolymer of dimethyl diallyl ammonium chloride and acrylic acid (550, Chemviron), poly- amino polyamide and their crosslinked water-soluble polymer, cation chitin Derivative, such as, for example, optionally in the quaternized chitosan of crystallite distribution, such as the dihalo- of dibromobutane etc. The condensation product of substituted alkyl and the double-dialkylamine of such as Bis-dimethylamino -1,3- propane etc., cationic guar gum are all Such as, for example, CelaneseCBS、C-17、C-16, QAS polymer, such as, example Such as, MiranolA-15、AD-1、AZ-1.Suitable anionic, amphion Type and amphoteric and non-ionic polyalcohol are, for example, vinylacetate/crotonic acid-copolymers, vinyl pyrrolidone/ Vinyl acrylate copolymer, vinylacetate/butyl maleate/iso-bornyl acrylate copolymer, methyl second Alkene ether/copolymer-maleic anhydride and its ester, uncrosslinked and polyol crosslink polyacrylic acid, acrylamidopropyl trimethyl Ammonium chloride/acrylate copolymer, octyl acrylamide/methyl methacrylate/tert-butyl group aminoethyl methacrylate/2- Hydroxypropyl methacrylate copolymer, polyvinylpyrrolidone, vinyl pyrrolidone/vinyl acetate copolymer, Vinyl pyrrolidone/dimethyl amino ethyl methacrylate/caprolactam terpolymer spreads out with optional Raw cellulose ether and organosilicon.

Pearlescent waxes

Suitable pearlescent waxes are：For example, alkylidene diol ester, particularly glycol distearate；Marlamid, Particularly fatty acid distribution of coconut oil diglycollic amide；Partial glyceride, particularly glyceryl monostearate；Polybasic ester, is optionally used The long-chain ester of the carboxylic acid, particularly tartaric acid of aliphatic alcohol hydroxyl substitution containing 6 to 22 carbon atoms；Aliphatic compound, it is all Such as, for example, fatty alcohol, aliphatic ketone, fatty aldehyde, aliphatic ether, and the aliphatic carbonate ester of at least 24 carbon atoms altogether is included, it is special It is not laurone (laurone) and distearyl acyl group ether；Aliphatic acid, such as stearic acid, hydroxy stearic acid or behenic acid, containing 12 to 22 The olefin epoxide of a carbon atom and the fatty alcohol containing 12 to 22 carbon atoms and/or contain 2 to 15 carbon atoms and 2 to 10 The open loop product and their mixture of the polyalcohol of a hydroxyl group.

Organosilicon

Suitable organo-silicon compound are for example, dimethyl polysiloxane, methyl phenyl silicone, cyclic organic and ammonia Base-, aliphatic acid-, alcohol-, polyethers-, epoxy-, fluoro-, glycocide-, and/or alkyl-modified polysiloxane compound, it is in room Can be both liquid and resinae under temperature.Other suitable polysiloxane compounds are dimethiones, it is that have to put down The dimethyl siloxane of a length of 200 to 300 dimethyl siloxane units of equal chain and the mixture of hydrogenated silicate.Suitably wave Being discussed in detail for organosilicon of hair property can be found in the Cosm.Toil.91,27 such as Todd (1976).

Wax and stabilizer

Except the natural oil used, wax is there may also be in preparation, more particularly native paraffin：Such as, for example, candelila wax, Brazil wax, Japan tallow, esparto wax (espartograss wax), cork wax, guaruma waxes, rice oil wax, sugarcane Wax, ouricury wax, lignite wax, beeswax, shellac wax, spermaceti, lanolin (lanolin) (lanocerin), tail fat are fatty, pure white Ceresine, ceresine (ceresine), vaseline, paraffin and microwax (microwaxes)；Chemical modification wax (hard wax), such as, for example, brown Coal ester type waxes, husky rope wax, hydrogenation Jojoba wax, and synthetic wax, such as, for example, polyalkylene wax and polyethylene glycol wax.Aliphatic acid Metal salt, such as, for example, ricinoleic acid or stearic magnesium salts, aluminium salt and/or zinc salt, are used as stabilizer.

Biological agent

In the context of the present invention, biological agent is, for example, tocopherol, tocopherol acetate, tocopheryl palmitate, anti- Bad hematic acid, (deoxidation) ribonucleic acid and its fragmentation products, beta glucan, retinol, bisabolol, allantoin, phytantriol, Panthenol, AHA acid, amino acid, ceramide, Questamide H, essential oil, plant extracts, for example, prune extract, class bar Draw berry extract and vitamin complex.

Film forming agent

The film forming agent of standard is, for example, chitosan, Microcrystalline chitosan, n-trimethyl chitosan chloride, polyvinylpyrrolidone, ethene Base pyrrolidones/vinyl acetate copolymer, the polymer of oleic series, quaternized cellulose (quaternary Cellulose derivatives), the salt and similar compound of collagen, hyaluronic acid with it.

Dandruff removing agent

Suitable dandruff removing agent is Pirocton Olamin (1- hydroxy-4-methyls -6- (2,4,4- trimethyl-pentyls) -2- (1H)-pyridone monoethanolamine salt),(climbazole),(4- acetyl -1- { 4- [2- (2,4- Dichlorophenyl) r-2- (1H- imidazoles -1- ylmethyls) -1,3- dioxane hex- c-4- ylmethoxies-phenyl piperazine, ketoconazole, Dichlorophenyl imidazoles dioxolanes (elubiol), selenium disulfide, sulikol, sulphur polyethylene glycol dehydrated sorbitol mono-fatty acid ester, The numb alcohol polyethoxylate of sulphur comb, sulphur tar distillate, salicylic acid (or composition with hexachlorophene), undecenoic acid, Dan Yi Alkylolamides sulfosuccinic acid sodium salt,UD (protein/undecenoic acid condensation product), zinc pyrithione, pyridine sulphur The pyrithione magnesium sulfate of ketone aluminium and pyrithione magnesium/bis-.

Preservative

Suitable preservative is, for example, phenoxetol, formalin, parabens, pentanediol or sorbic acid and The annex 6 in Kosmetikverordnung (" cosmetics regulations (Cosmetics Directive) "), is listed in part A and B Other classes compound.

Aromatic oil

Suitable aromatic oil is natural and synthetic perfume mixture.Natural perfume material includes flower (lily, lavender, rose, jasmine Jasmine, flores aurantii, Yilan) extract, stem and leaf (fish pelargonium, Pogostemon cablin, Petitgrain) extract, fruit (fennel, coriander, caraway, Du Pine) extract, pericarp (bergamot, lemon, orange) extract, root (nutmeg, Angelica sinensis, celery, cardamom, costus root, kite Tail, calamus (calmus)) extract, trees (pine, deodar, sandalwood, guaiaci lignum, narra) extract, grass and grass family (tarragon, lemon grass (Cymbopogon citratus), Salvia japonica, thyme) extract, pin and branch (dragon spruce, fir, pine tree, pinon pine) extract, resin and perfume (or spice) Cream (galbanum, elemi, styrax, myrrh, frankincense, hercules' allheal) extract.Animal material can also be used, for example, civetta and Castor fur.Typical synthetic perfume compound is the product of ester, ether, aldehyde, ketone, alcohol, hydro carbons.The example of the flavor compounds of esters Son is benzyl acetate, phenoxyethyl isobutyrate, p- t-butylcyclohexyl yl acetate, bergamio, dimethyl benzyl original Ester, phenethyl acetate, linalyl benzoate, benzyl formate, glycine ethyl methyl phenyl ester (ethyl methyl phenyl Glycinate), allyl cyclohexyl propionate, styralyl propionate and benzyl salicylate.Ethers may include, for example, benzyl ethyl Ether, and aldehyde includes, for example, linear alkanals, citral, citronellal containing 8 to 18 carbon atoms, lemongrass base oxygen acetaldehyde, cyclamen Aldehyde, laurine, lilial and bougeonal.The example of suitable ketone be irisone, isomethylionone, And vertofix coeur.Suitable alcohol is anethole, citronellol, eugenol, isoeugenol, geraniol, linalool, benzyl carbinol And terpineol.Hydrocarbon mainly includes terpenes and face cream.It is however preferred to use the mixture of different flavor compounds, Pleasant fragrance is produced together.Other suitable perfumery oils be with relative to low volatility use mostly as it is fragrant into The essential oil divided.Example is sage oil, Chamomile oil, caryophyllus oil, Mei Lisha oil, peppermint oil, cinnamon leaves oil, lime caul-fat, needle juniper Seed oil, vetiver oil, frankincense oil, maple seed oil, ladanum oil and lavender oil.It is preferably in the form of independent or mixture below The compound used：Bergamot oil, dihydromyrcenol lilial, lilial, lyral, citronellol, phenethyl alcohol, hexyl Cinnamic acid, geraniol, benzylacetone, cyclamen aldehyde, linalool, (ethoxymethyl) epoxide ring hendecane (Boisambrene), imperial saliva Furans (Ambroxan), indoles, dihydrojasmonate (hedione), 2- methyl -4- (2,2,3- trimethyl -3- rings penta Alkene -1- bases) -2- butene-1-ols (sandelice), citrus (Citrus) oil, tangerine (Mandarin) oil, orange oil, the fluffy ester of lattice, 3, 6- dimethyl -3- cyclohexenes-formaldehyde (cyclovertal), lavender oil, sage oil, damascone (damascene), bourbon Pelargonium oil, salicylic acid cyclohexyl, vertofix coeur, ambrotone, tonalid (fixolide NP), oak moss (evernyl), Iraldein gamma, phenylacetic acid, geranyl acetate, benzyl acetate, rose oxide, Romillat, irotyl, tert-butyl group ring Hexyl ethyl carbonate (Floramat).

Dyestuff

Suitable dyestuff be it is any suitable and and ratify the material for cosmetics purpose, for example, in Chemie publishing houses, Publication " the Kosmetische of the Farbstoff commission Deutschen researchs federation of WeinheimListed in 1984, page 81 to page 106.Example includes alkermes A (C.I.16255), patent Blue V (C.I.42051), indigo (C.I.73015), chlorophyllin (C.I.75810), quinoline yellow (C.I.47005), titanium dioxide (C.I.77891), indanthrene blue RS (C.I.69800) and madder red (C.I.58000).Luminol can also be used as luminescent dye Occur.Under normal circumstances, according to mixture as a whole, the concentration that these dyestuffs use is 0.001 to 0.1 weight %.

According to specific composition, the percent of total content of auxiliary agent and additive can from 1 to 50% (weight), be preferably 5 to 40% (weight).Composition can be produced by the hot or cold technique of standard.

Capsule and microcapsules

For oral, the encapsulation of extract represents preferred embodiment.Usually encapsulation can be by the use of gelatin as matrix come real Apply.Also can be by adding gelling agent (such as, such as alginate) to extract and obtained mixture instillation calcium salt bath In prepare capsule.Both approaches all obtain having a diameter of large capsule from about 1 centimetre to about 5 centimetres, it is toxicology Safety, and it is adapted to digestion.

The extract encapsulation of composite preparation for being developed for topical application is also likely to be needs.This can have different originals Cause：Resistance and the stabilization that other compounds interact in the formulation, prevent chemical degradation or non-simply for preparing The product of Chang Meiguan.For this purpose, commonly used microcapsules." microcapsules " are understood to spheroidal aggravation, its is a diameter of From about 0.1 to about 5 millimeter, include the core of at least one solid or liquid surrounded by least one continuous film.More precisely, They are finely divided liquid or scribble the solid of film forming polymer, in its production, emulsify and agglomerate or interfacial polymerization after, Polymer deposits are on material to be packaged.During another, liquid actives are absorbed in matrix (" microsponge "), And as the particulate that can be additionally coated with film forming polymer.Microcosmic Caplet, also referred to as Nano capsule, can be with same with powder Mode dry.Except single-core microcapsules, also there is multinuclear aggregation, be referred to as microballoon, it includes be distributed in continuous membrane material In two or more cores.In addition, the microcapsules of single or multiple core are surrounded by second, third extra film etc..The film It can be made of material that is natural, semi-synthetic or synthesizing.Natural membrane material has, for example, Arabic gum, agar, agarose, Maltodextrin, alginic acid, and it is their salt (such as mosanom or calcium salt), fat and aliphatic acid, cetanol, collagen, de- Chitosan, lecithin, gelatin, albumin, shellac, polysaccharide (such as starch or glucan), polypeptide, protein hydrolysate, Sucrose and wax.Semi-synthetic membrane material is the cellulose of chemical modification, more particularly cellulose esters and ether among other things, for example, Cellulose acetate, ethyl cellulose, hydroxypropyl cellulose, hydroxypropyl methyl cellulose and carboxymethyl cellulose and starch derive The ether and ester of thing, more particularly starch.Synthesis membrane material is, for example, polymer, such as polyacrylate, polyamide, polyvinyl alcohol Or polyvinylpyrrolidone.The example of known microcapsules is following commercial product (membrane material is shown in bracket)： Hallcrest microcapsules (gelatin, gum arabic), Coletica Thalaspheres (marine collagen), Lipotec Millicapseln (alginic acid, agar), Induchem Unispheres (lactose, microcrystalline cellulose, hydroxypropyl methyl fiber Element), Unicetin C30 (lactose, microcrystalline cellulose, hydroxypropyl methyl cellulose), Kobo Glycospheres it is (modified to form sediment Powder, fatty acid ester, phosphatide), Softspheres (modified agar) and Kuhs Probiol Nanospheres (phosphatide).

Non-drug application

In addition, the present invention also relates to numerous applications, more particularly to be selected from

(i) garlic Trentepohlia,

(ii) Thalassiosira,

(iii) Chaetoceros category, and/or

(iv) Chlorococcum,

Microalgae extract purposes, its

■ is used for the treatment of people's hair；

■ is used for the treatment of application on human skin；

■ is used to adjust people's hair and/or the melanogenesis of application on human skin；

■ is used for the growth of people's hair and hair follicle；

■ is used for the unnecessary growth for suppressing people's hair；

■ is used to resist and pre-Anti-hair loss；

■ is used to improve and stimulates the synthesis of the collagen in human dermis' layer；

■ is used to prevent and to resisting age of skin；

■ is used to improve and stimulates in application on human skin, the glucosaminoglycan synthesis especially in corium or epidermis；

■ is used to improve and stimulate the Keratinocyte differentiation in people's epidermis, for adjusting the cuticula in people's epidermis；

■ is used to improve and stimulates corium and the propagation of epidermal cell and wound healing；

■ is used to improve and stimulate melanocyte proliferation；

■ is used to improve and stimulate lipolysis；

It is people's skin that all these applications, which can summarize statement, particularly corium, and people's hair, particularly " the tune of hair follicle Section ", to resist or prevent the symptom of such as alopecia or trichochromes depigmentation etc or such as the one of papule or inflammation etc The relevant skin disease of dysfunction of a little hair follicles.

Hereinafter, by-but be not limited to-various embodiments illustrate the present invention.

Embodiment

A. introduction

In order to solve the problems, such as the basic complexity of the present invention, and assess the biological characteristics of microalgae component, it is necessary to logical Cross and prepare complementary extract and each complementary extract is carried out the specific filler test for being adapted to its prominent bioactivity, To separate its biomass.There is huge bio-diversity in view of microalgae, it should be readily understood that a large amount of related Work be it is necessary, so as to identify be adapted to provide for showing needed for performance extract only a few strain.Claimed original The preparation of material extract is a kind of simple but effective method, the relevant algae of compatibility of the Extraction solvent to separate and use The basis of biomass.The theme of this technology teaching is that the biology that some are innovated present in the composition of the species of consideration is living The displaying of property.How following experiment shows can obtain different bioactive extracts from identical algal biomass, and How they can handle biomass so as to continuously extract by being subsequently exposed to different solvents.

For the bioactivity of the example of the open microalgae studied, with from

■ garlics Trentepohlia (true eyespot algae guiding principle),

■ Thalassiosiras (Bacillariophyceae),

■ Chaetoceros category (Bacillariophyceae), and

■ Chlorococcums (Chlorococcum guiding principle)

Drying and powder biomass obtain different extracts carried out various experiments.

Experimental program is extracted from a lot of other selection of technical scheme, and they have been considered as certain example and represent. According to the present invention, liquid extraction agent of the cellular material of above-mentioned microalgae selected from ethyl acetate, isopropanol, ethanol, first alcohol and water To extract.

Extractant can include the mixture of two or more above-mentioned solvent.Hereinafter, the concentration of extract The ratio that can be routinely expressed as between the quantity (weight) of cell material and the quantity (volume) of Extraction solvent of processing.Example Such as, 1 gram of dry powder microalgae is handled by using 200 milliliters of Extraction solvents, obtains 200 milliliters of 5000 mcg/mls (weight/volume) Extract, without considering the quantity of the compound really dissolved in the solvent.This normal concentration, which allows to represent to produce, to be retouched The actually required microalgae quantity of the experimental result stated.However, table 1 reports the dry weight of the estimation of extract, and can calculate Go out real extract concentrations.Relative to cultural method and environmental condition, the composition of microalgae may change, and extraction efficiency can It can change, it is necessary to consider the dry weight of extract as original instruction.

The quality and quantity of compound present in extract is relative to both the property of solvent and the experimental program of preparation May be different.Selected algae strain is shown under the action of the solvent essentially according to the characteristic of their own cell membrane There is different resistance to degree (refractoriness) to h substance.The dry weight of the extract of preparation is reported in table 1, it is represented For the percentage of related overall microalgae material：

Table 1

The dry weight of extract, is expressed as the percentage of dry cellular material

Extract code	Chaetoceros category	Thalassiosira pseudonana	Garlic algae	Chlorococcum
					Methanol	46-58	39	19.5	18-28
Ethanol	32	27		6.5
					Isopropanol	22	15	2	3
Ethyl acetate	13-18	18	4	3-4
					Ethanol ＜ ethyl acetate	22	18	5	5
Water ＜ ethanol ＜ ethyl acetate	34	22	16	34-44
					Extract code	Calcareous Chaetoceros			Small Chlorococcum
Methanol	54%			24%
					Ethanol	22%			16%
Isopropanol	10%			10%
					Ethyl acetate	9-12%			5-8%
Ethanol ＜ ethyl acetate	13%			8-12%
					Water ＜ ethanol ＜ ethyl acetate	64%			22%

B. activity of the microalgae extract to hair follicle growth

Embodiment 1 to 2

Belong to activity of the direct methanol (d-MeOH) to hair follicle growth of extraction from Chaetoceros

Hair follicle is derived from the scalp sample of single contributor and is transferred to sterile 24 orifice plate with by using William's E culture mediums of improvement (Williams'Medium E) is cultivated.Culture carries out ten days, and the experiment process of hair follicle is small from the initial progress 24 of culture When.Hair follicle of the selection for experiment after when culture 18 is small.Only those show good life state and increase not low It is deemed suitable in 0.2 millimeter of hair follicle for culture.All experimental groups and control group prepared includes 12 hair follicles, 24 orifice plates are inoculated in the density in 3 hair follicles/hole.Caused presentation is substantially held due to non-experiment process in incubation The hair follicle for bearing (sufferance) sign is excluded from final analysis.In obtaining for the dried biomass that belongs to from Chaetoceros In the case of direct methanol extract (d-MeOH), following experiment is carried out to prove direct methanol extract (d-MeOH) to hair follicle The activity of growth.MeOH extracts prepared by the cell material that assay medium is done by Chaetoceros supplement, and correspond to 0.1 to obtain With the most latter two concentration of 1 mcg/ml (=ppm).The growth performance observed in the hair follicle of processing is not with adding The control group contrast cultivated in the identical culture medium of extract.The activity of microalgae processing is demonstrate,proved by the increase of hair follicle growth Real, the increase of hair follicle growth shows as the change of the average elongation of each experimental group compared with the control group.In the culture of nine days Experiment is terminated after (8 processing).Hair follicle grows through microphotograph to study, and is then determined by graphical analysis.It is all Culture of the hair follicle respectively the 5th day, the 9th day is taken photo.Experiment 3 times is repeated using the hair follicle for being derived from three donors.Report The data in road are calculated by the data that the hair follicle collected from the donor of response records, still, it was recently reported that the donor of response Quantity (being shown in Table 2).

Table 2

Growth-elongation of hair follicle is expressed as the % ratios of the performance of control group.Collect from two in the donor of three bit tests Data (response=66%)

Embodiment	Sample	Quantity	It is average	Standard error	The total quantity of hair follicle
						0	Control	0	100.00	3.09	36
1	d-MeOH	0.1μg/ml	103.18	4.82	22
						2	d-MeOH	1.0μg/ml	110.30	3.63	21

The result shows that addition direct methanol extract causes hair follicle growth to increase, compared with untreated group, become from 3% to 10% Change.Best response is obtained by using the extract-treated hair follicle of 1 mcg/ml.

Embodiment 3 to 4

Activity from direct ethanol (d-EtOH) extract that the short modification of calcareous Chaetoceros obtains to hair follicle growth

Previously described experimental program is used to study the direct ethanol extract obtained from the short modification of calcareous Chaetoceros to hair follicle The activity of growth.Experiment is repeated using the hair follicle for being derived from two donors twice.Report is calculated in data by collecting record The result in road and shown in table 3.

Table 3

Growth-elongation of hair follicle is expressed as the % ratios of control group performance.Collect from two in the donor of two bit tests Data (response=100%)

Embodiment	Sample	Quantity	It is average	Standard error	The total quantity of hair follicle	ANOVA
							0	Control	0	100,0	3.3	27	-
3	d-EtOH	0.1μg/ml	116,7	7.4	19.0	P ＜ 0.05
							4	d-EtOH	1μg/ml	119,5	8.4	16.0	P ＜ 0.05

Direct ethanol extract hair follicle stimulating growth, induces the increase of elongation, from 17% to 20% change.Both results have It is statistically significant, however, detecting best response by using the extract-treated hair follicle of 1 mcg/ml.

Embodiment 5 to 6

Activity from direct isopropyl (d-iPrOH) alcohol extracting thing that Chaetoceros category obtains to hair follicle growth

Previously described experimental program is used to study life of the direct isopropyl alcohol extract obtained from Chaetoceros category to hair follicle Long activity.In table 4, the result of single experiment is reported.

Table 4

Growth-elongation of hair follicle is expressed as the % ratios of control group performance.(data of single experiment)

Embodiment	Sample	Quantity	It is average	Standard error	The total quantity of hair follicle	ANOVA
							0	Control	0	100.0	4.9	14	-
5	d-iPrOH	0.1μg/ml	75.7	4.6	10	P ＜ 0.05
							6	d-iPrOH	10.0μg/ml	69.1	3.6	10	P ＜ 0.01

Direct isopropyl alcohol extracting thing suppresses hair follicle growth, causes the reduction of elongation, from -24% to -31% change.By using 10 The extract-treated hair follicle of mcg/ml detects best response, and the result has the statistical significance of highly significant.

Embodiment 7 to 9

The activity of growth of direct ethyl acetate (d-EtAc) extract obtained from Chaetoceros category to hair follicle

Previously described experimental program is used to study the direct ethyl acetate extract obtained from Chaetoceros category to hair follicle growth Activity.The data of report are calculated by the data that the hair follicle collected from the donor of response records, but, it was recently reported that contribute The quantity (being shown in Table 5) of person.

Table 5

Growth-elongation of hair follicle is expressed as the % ratios of the performance of control group.Collect from 3 in the donor of 5 bit tests Data (response=60%)

Embodiment	Sample	Quantity	It is average	Standard error	The total quantity of hair follicle	ANOVA
							0	Control	0	100.0	2.2	52
7	d-EtAc	0.1μg/ml	111.9	3.4	34	P ＜ 0.01
							8	d-EtAc	1.0μg/ml	110.2	3.5	36	P ＜ 0.05
9	d-EtAc	10.0μg/ml	112.4	3.3	35	P ＜ 0.01

All tests concentration direct ethyl acetate extract hair follicle stimulating growth, cause elongation to increase, from 10% to 12% change.Statistical significance of all responses with statistical significance or highly significant.

Embodiment 10 to 11

Activity from direct ethyl acetate (d-EtAc) extract that the short modification of calcareous Chaetoceros obtains to hair follicle growth

Previously described experimental program is used to study the extract of the direct ethyl acetate obtained from the short modification of calcareous Chaetoceros To the activity of hair follicle growth.The data (being shown in Table 6) of report are obtained from single donor.

Table 6

Growth-elongation of hair follicle is expressed as the % ratios of control group performance.(data of single experiment)

Embodiment	Sample	Quantity	It is average	Standard error	The total quantity of hair follicle
						0	Control	0	100.0	5.0	13
10	d-EtAc	0.1μg/ml	100.1	6.0	9
						11	d-EtAc	1.0μg/ml	110.6	7.8	9

Grown in the direct acetic acid ethyl acetate extract hair follicle stimulating of 1 mcg/ml, cause elongation to increase by 11%.As a result confirm The bioactivity of the extract obtained from the short modification of calcareous Chaetoceros, and the property of the extract with previously belonging to screening from Chaetoceros Unanimously.

Embodiment 12 to 13

The activity of growth of continuous ethanol (s-EtOH) extract obtained from Chaetoceros category to hair follicle

Previously described experimental program is used to study growth of the continuous ethanol extract obtained from Chaetoceros category to hair follicle Activity.Experiment four times is repeated using the hair follicle for being derived from four donors.The data of report are by collecting from all donors' The data of all hair follicles record calculate and (are shown in Table 7).

Table 7

Growth-elongation of hair follicle is expressed as the % ratios of the performance of control group.Collect number from 4 in the donor of 4 bit tests According to (response=100%)

Embodiment	Sample	Quantity	It is average	Standard error	The total quantity of hair follicle	ANOVA
							0	Control	0	100.0	2.3	70
12	s-EtOH	0.1μg/ml	104.2	3.0	46	Non-stimulated (n.s.)
							13	s-EtOH	1.0μg/ml	111.9	3.5	43	P ＜ 0.01

It is preferred that the continuous ethanol extract hair follicle stimulating in 1 mcg/ml is grown, the average amplification for causing elongation is 12%, The result has the statistical significance of highly significant.

Embodiment 14 to 15

Activity from continuous ethanol (s-EtOH) extract that the short modification of calcareous Chaetoceros obtains to hair follicle growth

Use experimental program noted earlier and hair follicle is given birth to studying from the continuous ethanol extract that the short modification of calcareous Chaetoceros obtains Long activity.The data (being shown in Table 8) of report are obtained from single donor.

Table 8

Growth-elongation of hair follicle is expressed as the % ratios of control group performance.

It is preferred that the continuous ethanol extract hair follicle stimulating in 1 mcg/ml is grown, the average amplification for causing elongation is 18%, The result has statistical significance.

As a result extract of the bioactivity for the extract for proving to obtain from the short modification of calcareous Chaetoceros with belonging to screening from Chaetoceros Property it is consistent.

Embodiment 16 to 17

Activity of direct methanol (d-MeOH) extract obtained from Thalassiosira to hair follicle growth

Experimental program noted earlier is used to study the work of growth of the direct methanol extract obtained from Thalassiosira to hair follicle Property.Experiment 3 times is repeated using the hair follicle for being derived from three donors.The data of report are by collecting the hair follicle from the donor of response The data of record calculate, still, it was recently reported that the quantity (being shown in Table 9) of donor.

Table 9

Growth-elongation of hair follicle is expressed as the % ratios of the performance of control group.Collect from 2 in the donor of 3 bit tests Data (response=66%)

Embodiment	Sample	Quantity	It is average	Standard error	The total quantity of hair follicle	ANOVA
							0	Control	0	100.0	3.1	36
16	d-MeOH	0.1μg/ml	111.6	4.6	21	P ＜ 0.05
							17	d-MeOH	1.0μg/ml	97.9	3.7	23	It is non-stimulated

Direct methanol extract hair follicle stimulating is grown, compared with control group, the elongation increase by 12% of generation.With 0.1 microgram/milli The extract-treated hair follicle risen obtains best response, this result has statistical significance.

Embodiment 18 to 20

Activity from the direct isopropyl alcohol extracting thing (d-iPrOH) that Thalassiosira obtains to hair follicle growth

Experimental program noted earlier is used to study from Thalassiosira pseudonana (Thalassiosira pseudonana) acquisition The activity of direct growth of the isopropyl alcohol extracting thing to hair follicle.Single experiment (table 10) is carried out.

Table 10

Growth-elongation of hair follicle is expressed as the % ratios of the performance of control group.(from single experimental data)

Embodiment	Sample	Quantity	It is average	Standard error	The total quantity of hair follicle	ANOVA
							0	Control	0	100.0	4.9	14
18	d-iPrOH	0.1μg/ml	77.5	8.0	9	P ＜ 0.05
							19	d-iProH	1.0μg/ml	85.1	8.2	10	It is non-stimulated
20	d-iPrOH	10.0μg/ml	87.3	4.2	10	It is non-stimulated

Direct isopropyl alcohol extracting thing suppresses the growth of hair follicle, causes elongation to reduce, from -12% to -22% change.It is preferred that 1 The extract-treated hair follicle of mcg/ml obtains best response, this result has statistical significance.

Embodiment 21 to 23

Activity from direct ethyl acetate (d-EtAc) extract that Thalassiosira obtains to hair follicle growth

Use experimental program noted earlier and hair follicle is given birth to studying from the direct ethyl acetate extract that Thalassiosira pseudonana obtains Long activity.Experiment 5 times is repeated using the hair follicle for being derived from five donors.The data of report are by collecting the donor from response The data of hair follicle record calculate, still, it was recently reported that the quantity (being shown in Table 11) of donor.

Table 11

Growth-elongation of hair follicle is expressed as the % ratios of the performance of control group.Collect from 4 in the donor of 5 bit tests Data (response=80%)

Embodiment	Sample	Quantity	It is average	Standard error	The total quantity of hair follicle	ANOVA
							0	Control	0	100	2.2	65
21	d-EtAc	0.1μg/ml	95.9	2.8	38	It is non-stimulated
							22	d-EtAc	1.0μg/ml	115.2	3.6	44	P ＜ 0.01
23	d-EtAc	10.0μg/ml	106.0	2.8	46	It is non-stimulated

It is preferred that the direct ethyl acetate extract hair follicle stimulating in 1 mcg/ml is grown, the average amplification for causing elongation is 15%, these results have the statistical significance of highly significant.

Embodiment 24 to 25

The activity of growth of continuous ethanol (s-EtOH) extract obtained from Thalassiosira to hair follicle

Experimental program noted earlier is used to study growth of the continuous ethanol extract obtained from Thalassiosira pseudonana to hair follicle Activity.Experiment 4 times is repeated using the hair follicle for being derived from four donors.The data of report are by collecting from the donor's of response The data of hair follicle record calculate, still, it was recently reported that the quantity (being shown in Table 12) of donor.

Table 12

Growth-elongation of hair follicle is expressed as the % ratios of the performance of control group.Collect from 3 in the donor of 4 bit tests Data (response=75%)

Embodiment	Sample	Quantity	It is average	Standard error	The total quantity of hair follicle	ANOVA
							0	Control	0	100.0	2.9	52	-
24	s-EtOH	0.1μg/ml	112.2	3.3	31	P ＜ 0.01
							25	s-EtOH	1.0μg/ml	110.5	3.7	28	P ＜ 0.05

Continuous ethanol extract hair follicle stimulating growth, compared with control group, causes elongation to increase, from 10% to 12% change. The result that both processing produce has statistical significance.Best sound is detected by using 0.1 mcg/ml processing hair follicle Should, the elongation of the generation amplification that be averaged is 12%, and which results in the statistical significance of highly significant.

Embodiment 26 to 27

Activity from the direct methanol extract (d-MeOH) that garlic Trentepohlia obtains to hair follicle growth

Experimental program noted earlier is used to study the activity from the direct methanol extract that garlic algae obtains to hair follicle growth.Profit Experiment 3 times is repeated with the hair follicle for being derived from three donors.The data of report are recorded by the hair follicle collected from the donor of response Data calculate, still, it was recently reported that the quantity (being shown in Table 13) of donor.

Table 13

Growth-elongation of hair follicle is expressed as the % ratios of the performance of control group.Collect from 2 in the donor of 3 bit tests Data (response=66%)

Direct methanol extract hair follicle stimulating is grown, and compared with control group, causes elongation to increase, from 15% to 18% change. The result that both processing produce has significant statistical significance；However, by using the processing hair follicle detection of 0.1 mcg/ml To best response, the elongation of generation is averaged amplification as 18%, as a result the statistical significance with highly significant.

Embodiment 28 to 30

Activity from the direct ethanol extract (d-EtOH) that garlic Trentepohlia obtains to hair follicle growth

Experimental program noted earlier is used to study the activity from the direct ethanol extract that garlic algae obtains to hair follicle growth.Point Do not tried by using three kinds of concentration for the treatment of processing hair follicle of 0.1 mcg/ml, 1.0 mcg/mls, 10.0 mcg/mls Test, experimental result is shown in Table 14.

Table 14

One elongation of growth of hair follicle is expressed as the % ratios of the performance of control group (from single experimental data)

Embodiment	Sample	Quantity	It is average	Standard error	The total quantity of hair follicle	ANOVA
							0	Control	0	100	4.7	18
28	d-EtOH	0.1μg/ml	99.8	3.7	11	It is non-stimulated
							Z9	d-EtOH	1.0μg/ml	106.4	3.8	12	It is non-stimulated
30	d-EtOH	10.0μg/ml	125.0	6.8	11	P ＜ 0.001

As a result the extract proved in the concentration of 10 mcg/mls has strong impulse effect, compared with control group, generate 25% amplification.

Embodiment 31 to 33

Activity from direct ethyl acetate (d-EtAc) extract that garlic Trentepohlia obtains to hair follicle growth

Experimental program noted earlier is used to study work of the direct ethyl acetate extract obtained from garlic algae to hair follicle growth Property.Experiment 3 times is repeated using the hair follicle for being derived from three donors.The data of report are by collecting the hair follicle from the donor of response The data of record calculate, still, it was recently reported that the quantity (being shown in Table 15) of donor.

Table 15

Growth-elongation of hair follicle is expressed as the % ratios of the performance of control group.Collect from 3 in the donor of 3 bit tests Data (response=100%)

Embodiment	Sample	Quantity	It is average	Standard error	The total quantity of hair follicle	ANOVA
							0	Control	0	100.0	2.3	49
31	d-EtAc	0.1μg/ml	111.1	3.9	34	P ＜ 0.05
							32	d-EtAc	1.0μg/ml	110.7	3.7	33	P ＜ 0.05
33	d-EtAc	10.0μg/ml	110.2	3.9	20	It is non-stimulated

Grown in the direct ethyl acetate extract of all concentration for the treatment of with similar intensity hair follicle stimulating, however, only existing The result of 0.1-1.0 mcg/mls has statistical significance, and the average amplification for causing elongation is 11%.

Embodiment 34 to 35

Activity from the continuous ethanol extract (s-EtOH) that garlic Trentepohlia obtains to hair follicle growth

Experimental program noted earlier is used to study the activity from the continuous ethanol extract that garlic algae obtains to hair follicle growth.Profit Experiment 2 times is repeated with the hair follicle for being derived from two donors.The data of report are recorded by the hair follicle collected from the donor of response Data calculate, still, it was recently reported that the quantity (being shown in Table 16) of donor.

Table 16

Growth-elongation of hair follicle is expressed as the % ratios of the performance of control group.Collect from 2 in the donor of 2 bit tests Data (response=100%)

Embodiment	Sample	Quantity	It is average	Standard error	The total quantity of hair follicle	ANOVA
							0	Control	0	100.0	2.9	33
34	s-EtOH	0.1μg/ml	118.1	4.4	24	P ＜ 0.01
							35	s-EtOH	1.0μg/ml	115.2	5.6	22	P ＜ 0.05

Compared with control group, continuous ethanol extract hair follicle stimulating grows and produces elongation increase, from 15% to 18% change. The result that both processing tests produce has significant statistical significance.It is preferred that being handled in 0.1 mcg/ml, cause Elongation be averaged amplification as 18%, the result of generation has the statistical significance of highly significant.

Embodiment 36 to 37

Activity of direct methanol (d-MeOH) extract obtained from Chlorococcum to hair follicle growth

Experimental program noted earlier is used to study the activity from the direct methanol extract that Chlorococcum obtains to hair follicle growth. Experiment 3 times is repeated using the hair follicle for being derived from three donors.The data of report are remembered by the hair follicle collected from the donor of response The data of record calculate, still, it was recently reported that the quantity (being shown in Table 17) of donor.

Table 17

Growth-elongation of hair follicle is expressed as the % ratios of the performance of control group.Collect from 3 in the donor of 3 bit tests Data (response=100%)

Embodiment	Sample	Quantity	It is average	Standard error	The total quantity of hair follicle	ANOVA
							0	Control	0	100.0	1.8	50
36	d-MeOH	0.1μg/ml	109.5	3.7	32	P ＜ 0.05
							37	d-MeOH	1.0μg/ml	108.5	3.4	33	P ＜ 0.05

Compared with control group, the growth of direct methanol extract hair follicle stimulating, produces elongation increase, becomes from 8.5% to 9.5% Change.All results have statistical significance；But detect best response in 0.1 mcg/ml processing hair follicle.

Embodiment 38 to 39

Activity from the direct ethanol extract (d-EtOH) that Chlorococcum obtains to hair follicle growth

Experimental program noted earlier is used to study the activity from the direct ethanol extract that Chlorococcum obtains to hair follicle growth. Tested respectively with two kinds of concentration for the treatment of, 0.1 mcg/ml and 1 mcg/ml processing hair follicle.As a result it is reported in table 18 In.

Table 18

Growth-elongation of hair follicle is expressed as the % ratios of the performance of control group.(coming from single experimental data)

Embodiment	Sample	Quantity	It is average	Standard error	The total quantity of hair follicle	ANOVA
							0	Control	0	100.0	4.7	18
38	d-EtOH	0.1μg/ml	109.6	6.4	11	It is non-stimulated
							39	d-EtOH	1.0μg/ml	118.8	4.5	11	P ＜ 0.05

The results show that compared with control group, strong impulse effect is produced in the extract of the concentration of 1.0 mcg/mls, is produced Growth increase by 19%.The response has statistical significance (P ＜ 0.05).

Embodiment 40 to 42

Activity from direct isopropanol (d-iPrOH) extract that Chlorococcum obtains to hair follicle growth

Experimental program noted earlier is used to study work of the direct isopropyl alcohol extracting thing obtained from Chlorococcum to hair follicle growth Property.Data in relation to reporting obtain (table 19) from single experiment.

Table 19

Growth-elongation of hair follicle is expressed as the % ratios of the performance of control group.Data from single donor

Embodiment	Sample	Quantity	It is average	Standard error	The total quantity of hair follicle	ANOVA
							O	Control	0	100.0	4.9	14
40	d-iPrOH	0.1μg/ml	94.0	7.3	10	It is non-stimulated
							41	d-iPrOH	1.0μg/ml	85.0	5.2	10	It is non-stimulated
42	d-IPrOH	10.0μg/ml	81.2	4.4	11	P ＜ 0.05

Compared with control group, direct isopropyl alcohol extracting thing suppresses the growth of hair follicle, produces the reduction from -6% to -19% of elongation Change.Best response is obtained by using the extract-treated hair follicle of 10 mcg/mls, and its result has statistical significance.

Embodiment 43 to 45

Activity from direct ethyl acetate (d-EtAc) extract that Chlorococcum obtains to hair follicle growth

Experimental program noted earlier is used to study the direct ethyl acetate extract obtained from Chlorococcum to hair follicle growth Activity.Experiment 3 times is repeated using the hair follicle for being derived from three donors.The data of report are by collecting the hair from the donor of response The data of capsule record calculate, and still, report the quantity (being shown in Table 20) of donor.

Table 20

Growth-elongation of hair follicle is expressed as the % ratios of the performance of control group.Collect from 3 in the donor of 3 bit tests Data (response=100%)

Embodiment	Sample	Quantity	It is average	Standard error	The total quantity of hair follicle	ANOVA
							0	Control	0	100.0	1.9	50
43	d-EtAc	0.1μg/ml	114.1	3.6	34	P ＜ 0.01
							44	d-EtAc	1.0μg/ml	105.8	3.1	34	It is non-stimulated
45	d-EtAc	10.0μg/ml	104.5	3.8	23	It is non-stimulated

Grown in the equal hair follicle stimulating of the direct ethyl acetate extract of all concentration for the treatment of, however, being examined in 0.1 mcg/ml Best response is measured, causes 14% growth amplification.The response has the statistical significance of highly significant.

Embodiment 46 to 47

Activity from direct ethyl acetate (d-EtAc) extract that small Chlorococcum obtains to hair follicle growth

Experimental program noted earlier is used to study the direct ethyl acetate extract obtained from small Chlorococcum to hair follicle growth Activity.The hair follicle that single donor is derived from by test obtains the result (table 21) reported.

Table 21

Growth-elongation of hair follicle is expressed as the % ratios (data from single experiment) of the performance of control group

Embodiment	Sample	Quantity	It is average	Standard error	The total quantity of hair follicle
						0	Control	0	100.0	5.0	13
46	d-EtAc	0.1μg/ml	109.7	5.4	11
						47	d-EtAc	1.0μg/ml	107.7	5.1	12

Compared with control group, direct ethyl acetate extract hair follicle stimulating growth, produces elongation increase, becomes from 8% to 10% Change.It is preferred that handled in 0.1 mcg/ml, elongation is caused to be averaged amplification as 10%.

As a result prove the bioactivity of extract that is obtained from small Chlorococcum with previously from the extract of Chlorococcum screening Property is consistent.

Embodiment 48 to 50

The continuous ethanol extract (S-EtOH ＜ EtAc) obtained from Chlorococcum is to the activity of hair follicle growth

Experimental program noted earlier is used to study the activity from the continuous ethanol extract that Chlorococcum obtains to hair follicle growth. Experiment is replicated using the hair follicle for being derived from two donors twice.The data of report are remembered by the hair follicle collected from the donor of response The data of record calculate, and still, report the quantity (being shown in Table 22) of donor.

Table 22

Growth-elongation of hair follicle is expressed as the % ratios of the performance of control group.Collect from 2 in the donor of 2 bit tests Data (response=100%)

Embodiment	Sample	Quantity	It is average	Standard error	The total quantity of hair follicle	ANOVA
							0	Control	0	100.0	2.3	34
48	s-EtOH	0.1μg/ml	118.5	4.3	22	P ＜ 0.001
							49	s-EtOH	1.0μg/ml	116.7	3.2	21	P ＜ 0.001
50	s-EtOH	10.0μg/ml	100.4	4.8	11	It is non-stimulated

Compared with control group, continuous ethanol extract hair follicle stimulating growth, the elongation increase of generation, becomes from 17% to 19% Change.System of the response with highly significant of processing generation is carried out in two kinds of concentration for the treatment of of 0.1 mcg/ml and 1 mcg/ml Meter learns meaning.It is preferred that being handled in 0.1 mcg/ml, the average amplification for causing elongation is 19%, this result has very Significant statistical significance.

B. to activity-overall conclusion of hair growth

The embodiment of the present invention confirms metabolic strong activity of the extract to hair follicle of research.Widely extracting Thing and direct extract, such as in direct methanol extract and in the extract continuously prepared, (it is included according to solvent polarity ladder Spend separated compound) in the case of, detect these activity.All results all prove to include in identical microalgae material Some active matters, are potentially suitable for producing opposite effect to hair follicle.In fact, extract activity is dependent on the preparation used Experimental program.Different solvents can be selected, to extract the compound with given activity.Prepared using different solvents continuous Extract shows that they include different active matters, although they are obtained (table 23) from identical microalgae cell material.

Table 23

To as caused by the extract studied to the disclosed active comprehensive analysis of the elongation of hair growth, elongation table It is shown as the % ratios (optimal average response) of the performance of control group

Extract code	d-MeOH	d-EtOH	d-iPrOH	d-EtAc	S-EtOH ＜ d-EtAc
						Chaetoceros category	+ 10%		- 24%	+ 12%	+ 12%
Calcareous Chaetoceros		+ 20%		+ 11%	+ 12% /+18%
						Thalassiosira pseudonana	+ 8%		- 22%	+ 15%	+ 12%
Garlic algae	+ 17%	+ 25%		+ 12%	+ 18%
						Chlorococcum	+ 10%	+ 19%	- 19%	+ 14%	+ 19%
Small Chlorococcum				+ 10%

C. for disclosed in the microalgae extract of consideration to the activity of melanogenesis

Melanocyte is responsible for the cell of the melanogenesis in skin and hair follicle.Melanin is the accumulation in hair and skin Pigment, and to be easy to quantitatively be adjusted in response to the exposure of sunlight, ageing process and last pathologic event.

Therefore, for body exterior in social life, and the healthy shape for effective holding skin and hair is presented State and youthful appearance state, the possibility for adjusting melanogenesis show dependent-chance in cosmetics.

Prepare excised human skin by screening, have studied the activity that microalgae extract generates melanin, to prove to whole group The effect knitted.

The experiment carried out on excised human skin's culture

The organ culture of Full thickness human skin is carried out, using dermatological specimens as starting, cutting diameter is about 7 millimeters of cylindrical shape sample And they are cultivated 6 days.The culture medium of use is the William-E of improvement, and the 3rd day in tissue cultures re-replaces. The sample of continuous extract is then dissolved in quantitative DMSO and is suitable for obtaining the final of 1 and 10 mcg/mls with being air-dried Concentration.The dermatological specimens of culture are applied topically to 4 microlitres these extract formulations daily.By the organ cultures of six days it Afterwards, histotomy is prepared from dermatological specimens, and uses quantifying for melanin content of Fontana-Masson Study on Technology of Dyeing Change.Melanin is obtained by the graphical analysis for the microphoto for cutting into slices to each artificial skin to quantify.

Embodiment 51 to 56

The activity that the continuous extract of three steps obtained from garlic Trentepohlia generates melanin

The continuous extract of three steps (ethyl acetate ＞ ethanol ＞ water) is prepared for from garlic algae.By as described above to mankind's skin Skin sample is handled, and screens these extracts.Experiment is repeated using the dermatological specimens for being derived from two donors twice.Pass through remittance Collect the data from two experiments and carry out result of calculation, as shown in table 24.

Table 24

The activity of melanogenesis in application on human skin of the continuous extract obtained from garlic Trentepohlia to cultured in vitro.Melanin content It is expressed as the % ratios of the performance of control group.The data (response=100%) collected from 2 in the donor of 2 bit tests

Embodiment	Sample	Quantity	It is average	Standard error	The quantity of sample	ANOVA
							0	Control	0	100.0	5.4	24
51	d-EtAc	1.0μg/ml	92.2	3.7	24	It is non-stimulated
							52	d-EtAc	10μg/ml	113.0	6.4	24	It is non-stimulated
53	s-EtOH	1.0μg/ml	75.3	3.7	24	P ＜ 0.01
							54	s-EtOH	10μg/ml	86.2	4.9	24	P ＜ 0.05
55	s-Water	1.0μg/ml	80.5	6.1	24	P ＜ 0.01
							56	s-Water	10μg/ml	101.2	5.3	24	It is non-stimulated

The results show that compared with control group, continuous ethanol extract and continuous water extract suppress melanogenesis, cause melanin Content reduces by from -19% to -25% change.As a result there is the statistical significance of statistical significance or highly significant.On the contrary, directly Ethyl acetate extract does not carry out the adjusting of any significant melanogenesis.

Embodiment 57 to 62

Activity from the continuous extract of three steps that Chlorococcum obtains to melanogenesis

The continuous extract of three steps (ethyl acetate ＞ ethanol ＞ water) is prepared from Chlorococcum.By as described above to human skin Sample is handled, and screens these extracts.Experiment is repeated using the dermatological specimens for being derived from two donors twice.By collecting Data from two experiments carry out result of calculation, as shown in Table 25.

Table 25

The activity of melanogenesis in application on human skin of the continuous extract obtained from Chlorococcum to cultured in vitro.Melanin content It is expressed as the % ratios of the performance of control group.The data (response=100%) collected from 2 in the donor of 2 bit tests

Embodiment	Sample	Quantity	It is average	Standard error	The quantity of sample	ANOVA
							0	Control	0	100.0	5.4	24
57	d-EtAc	1.0μg/ml	90.7	4.9	24	It is non-stimulated
							58	d-EtAc	10μg/ml	85.9	4.3	24	P ＜ 0.05
59	s-EtOH	1.0μg/ml	85.2	2.5	24	P ＜ 0.05
							60	s-EtOH	10μg/ml	80.8	3.4	24	P ＜ 0.01
61	s-Water	1.0μg/ml	102.5	5.5	24	It is non-stimulated
							62	s-Water	10μg/ml	99.9	4.4	24	It is non-stimulated

The results show that directly ethyl acetate extract and continuous ethanol extract suppress melanogenesis, cause melanin content with Control group is compared to reduction -8% to -19%.As a result there is the statistical significance of statistical significance or highly significant.It is on the contrary, continuous Water extract does not adjust melanogenesis.

Embodiment 63 to 74

Activity from the continuous extract of three steps that small Chlorococcum and the short modification of calcareous Chaetoceros obtain to melanogenesis

Respectively three steps continuous extract (ethyl acetate ＞ ethanol is prepared from small Chlorococcum (C) and the short modification of calcareous Chaetoceros (K) ＞ water).By handling application on human skin sample as described above, these extracts are screened.As a result it is as shown in table 26.

Table 26

In application on human skin of the continuous extract obtained from small Chlorococcum (C) and the short modification of calcareous Chaetoceros (K) to cultured in vitro Melanogenesis activity.Melanin content is expressed as the % ratios of the performance of control group.

Embodiment	Strain	Sample	Quantity	It is average	Standard error	The quantity of sample	ANOVA
								0		Control	0	100.0	7.3	21
63	C	d-EtAc	1.0μg/ml	105.9	5.6	10	It is non-stimulated
								64	C	d-EtAc	10μg/ml	95.6	6.1	10	It is non-stimulated
65	C	s-EtOH	1.0μg/ml	86.7	4.7	10	It is non-stimulated
								66	C	s-EtOH	10μg/ml	81.4	5.2	10	P ＜ 0.05
67	C	s-Water	1.0μg/ml	107.7	5.3	10	It is non-stimulated
								68	C	s-Water	10μg/ml	82.7	4.4	10	P ＜ 0.05
69	K	d-EtAc	1.0μg/ml	78.5	9.1	10	P ＜ 0.05
								70	K	d-EtAc	10μg/ml	78.2	4.5	10	P ＜ 0.05
71	K	s-EtOH	1.0μg/ml	78.0	5.1	10	P ＜ 0.05
								72	K	s-EtOH	10μgg/ml	69.7	3.4	10	P ＜ 0.01
73	K	s-Water	1.0μg/ml	88.2	9.9	10	It is non-stimulated
								74	K	s-Water	10μg/ml	79.3	6.5	10	P ＜ 0.05

The results show that compared with control group, obtain s-EtOH extracts from Chlorococcum and s- water extracts suppress melanogenesis, lead Blackening pigment content is reduced, from -13% to -19% change.There is statistics for the result of two kinds of processing in 10 mcg/mls Learn meaning.All extracts obtained from Chaetoceros generate relevant melanogenesis and suppress, compared with control group, from -12% To -30% change.These results have the statistical significance of statistical significance or highly significant；With the sEtOH of 10 mcg/mls Extract-treated skin, detects stronger response, it causes melanin content to reduce -30%.

Embodiment 75 to 82

The direct methanol extract obtained from small Chlorococcum, garlic algae, Thalassiosira pseudonana, the short modification of calcareous Chaetoceros is to black The activity of element generation

Prepared respectively from small Chlorococcum (C), garlic algae (M), Thalassiosira pseudonana (T) and the short modification of calcareous Chaetoceros (K) straight Connect methanolic extract (MeOH).By handling application on human skin sample as described above, these extracts are screened.As a result such as the institute of table 27 Show.

Table 27

The methanolic extract obtained from small Chlorococcum, garlic algae, Thalassiosira pseudonana and the short modification of calcareous Chaetoceros is in vitro The activity of melanogenesis in the application on human skin of culture.Melanin content is expressed as the % ratios of the performance of control group.

Embodiment	Strain	Sample	Quantity	It is average	Standard error	The quantity of sample	ANOVA
								0		Control	0	100.0	7.2	10
75	C	d-MeOH	1.0μg/ml	68.2	4.5	10	P ＜ 0.01
								76	C	d-MeOH	10μg/ml	81.4	7.6	10	It is non-stimulated
77	M	d-MeOH	1.0μg/ml	83.8	7.9	10	It is non-stimulated
								78	M	d-MeOH	10μg/ml	84.1	5.9	10	It is non-stimulated
79	T	d-MeOH	1.0μg/ml	74.6	9.0	10	P ＜ 0.05
								80	T	d-MeOH	10μg/ml	69.2	7.2	10	P ＜ 0.01
81	K	d-MeOH	1.0μg/ml	95.6	7.0	10	It is non-stimulated
								82	K	d-MeOH	10μg/ml	81.5	7.7	10	It is non-stimulated

The results show that compared with control group, all extracts suppress melanogenesis, cause melanin content to reduce, from -5% To -32% change.For with the processing of the 1 mcg/ml extract obtained from Chlorococcum (compared with control group, reduce- 32%) processing (compared with control group, reducing -31%) extract of 10 mcg/mls and from hailian seaweed obtained, the result Statistical significance with highly significant.

D. to activity of microalgae disclosed in primary Skin Cell

For microalgae extract to the activity from the separated primary fibroblast of application on human skin or horn cell, these microalgaes are screened Extract.The purpose of screening is the potential stimulating activity of synthesis of the investigation to I-type collagen and hyaluronic acid.Type i collagen Albumen is a kind of albumen, and hyaluronic acid is a kind of glucosaminoglycan, both is the important component of corium, due to skin The effect of aging, corium, which is subjected to significant quality and quantity, to be reduced.

The culture medium supplemented with microalgae extract is screened on primary cell culture, cell is closed with assessing microalgae Into the activity of collagen and hyaluronic acid.

Pass through the adjusting of primary skin fibroblasts research collagen synthesis

Experimentation is based on following steps：

■ primary fibroblasts are inoculated in 96 hole microwell plates, density for 20000 cells/centimetre²；

After when ■ cultures 24 are small, the culture medium of culture medium supplement is replaced, the culture substrate of supplement by 0.1 microgram of addition/ It is prepared by the extract obtained from above-mentioned microalgae strain of milliliter, 1.0 mcg/mls or 10 mcg/mls.Two serial octals are special Door is used for the screening of each culture medium through supplement.Keep two serial octals as a control group, and in standard medium Culture；

After when ■ cultures 48-72 is small, fibroblast is close and converges, and collagen directly on culture dish by carrying out Enzyme linked immunosorbent assay (ELISA) (ELISA) is quantified.The enzyme linked immunosorbent assay (ELISA) experimental program of use is exclusively for these realities Setting is tested, however, the upper similar experimental method of design reports (2007, BMC Cardiovascular by Jenkins etc. Disorders,7:13).For each treatment group, 8 holes carry out MTT experiment, to assess final cell density, and remaining 8 Hole carries out ELISA experiments, to assess the collagen of generation.Same operation is performed in control group.ELISA values divided by assessment Cell density, be assumed to be the quantitative index of the collagen of standardization.The data obtained from the group of processing are expressed as by right According to the percentage of the expressed value of group.

Embodiment 83 to 88

Direct methanol (d-MeOH) extract obtained from Chaetoceros category, garlic Trentepohlia and Chlorococcum is to Collagen of Fibroblasts The activity of albumen synthesis

Screen from Chaetoceros category (K), garlic algae (M) and Chlorococcum (C) acquisition d-MeOH extracts, with assess they to by The adjustment effect for the collagen synthesis that dermal fibroblast carries out.Processing primary fibroblast as described above, and can not Detect to precognition that the relevant collagen synthesis for responding the processing is adjusted.The results are shown in table 28.

Table 28 detects the work to corium by using the d-MeOH extract-treated primary fibroblasts of 0.1 and 10 microlitre/milliliter The amount of property-collagen is expressed as the % ratios of control group

Embodiment	Sample	Strain	Quantity	It is average	Standard error	The quantity in hole	ANOVA
								0	Control	-	0	100.0	1.7	8
83	d-MeOH	K	0.1μg/ml	135.9	10.7	7	P ＜ 0.01
								84	d-MeOH	K	10μg/ml	177.2	6.4	8	P ＜ 0.01
85	d-MeOH	M	0.1μg/ml	150.2	11.3	8	P ＜ 0.01
								86	d-MeOH	M	10μg/ml	158.2	14.1	8	P ＜ 0.01
87	d-MeOH	C	0.1μg/ml	161.9	11.2	8	P ＜ 0.01
								88	d-MeOH	C	10μg/ml	194.2	12.2	8	P ＜ 0.01

From the extract that Chaetoceros, garlic algae and Chlorococcum obtain all equal intense stimulus collagens of concentration for the treatment of conjunction Into.The result has the statistical significance of highly significant.

Embodiment 89 to 94

Direct ethanol (d-EtOH) extract and continuous water (s- water) extract obtained from Chaetoceros category is to fibroblast glue The activity of former albumen synthesis

Screening is by performing the direct ethanol extract and continuous water extract that the extraction of two steps is obtained from Chaetoceros category, to comment Estimate their adjustment effects to the synthesis of Collagen of Fibroblasts albumen.Trained according to previously described experimental program in fibroblast Support and screen extract on thing.As a result it is reported in table 29.

Table 29

By handling primary fibroblast, the % ratios that control group is expressed as to the amount of activity-collagen of corium are detected

Embodiment	Sample	Quantity	It is average	Standard error	The quantity in hole	ANOVA
							0	Control	0	100.0	3.9	8	-
89	d-EtOH	0.1μg/ml	163.4	4.3	8	P ＜ 0.01
							90	d-EtOH	1.0μg/ml	132.6	11.0	8	P ＜ 0.01
91	d-EtOH	10μg/ml	137.2	5.4	8	P ＜ 0.01
							92	s-Water	0.1μg/ml	143.5	8.9	8	P ＜ 0.01
93	s-Water	1.0μg/ml	115.6	7.5	8	It is non-stimulated
							94	s-Water	10μg/ml	128.9	6.1	8	P ＜ 0.01

The extract obtained from Chaetoceros, in almost all of concentration for the treatment of to the synthetically produced thorn strongly of collagen Swash effect.The result has the statistical significance of highly significant.

Embodiment 95 to 102

Direct ethyl acetate (d-EtAc) extract, the continuous ethanol (s-EtOH) obtained from the short modification of calcareous Chaetoceros extracts The activity that thing and continuous water (s-Water) extract synthesize Collagen of Fibroblasts albumen

The continuous extract of three steps (d-EtAc ＞ s-EtOH ＞ s-Water) obtained from the short modification of calcareous Chaetoceros is screened, to comment Estimate their adjustment effects to the synthesis of Collagen of Fibroblasts albumen.Trained according to previously described experimental program in fibroblast Support and screen extract on thing.As a result it is reported in table 30.

Table 30

By handling primary fibroblast, the % ratios that control group is expressed as to the amount of activity-collagen of corium are detected

Embodiment	Sample	Quantity	It is average	Standard error	The quantity in hole	ANOVA
							0	Control	0	100.0	3.5	8	-
95	d-EtAc	0.1μg/ml	107.5	1.9	8	It is non-stimulated
							96	d-EtAc	1.0μg/ml	105.7	6.2	8	It is non-stimulated
97	d-EtAc	10μg/ml	124.6	2.6	8	P ＜ 0.01
							98	s-EtOH	0.1μg/ml	100.6	12.0	8	It is non-stimulated
99	s-EtOH	1.0μg/ml	120.7	2.9	8	P ＜ 0.01
							100	s-Water	0.1μg/ml	112.2	3.2	8	It is non-stimulated
101	s-Water	1.0μg/ml	102.0	5.5	8	It is non-stimulated
							102	s-Water	10μg/ml	102.4	2.9	8	It is non-stimulated

The generation of all extract stimulation collagens, makes collagen synthesis increase by 12% (S- water extracts) to 25% (d- EtAc extracts).However, compared with control group, maximally related work is obtained by using the d-EtAc processing cells of 10 mcg/mls Property, it makes the synthesis of collagen add 25%, and with the s-EtOH processing of 1 mcg/ml, cause the collagen Synthesis increase by 21%.Both results have the statistical significance of highly significant (P ＜ are O.01).In the continuous of 0.1 mcg/ml The generation of water extract stimulation collagen is increased up to 12%.

Embodiment 103 to 108

Direct methanol (d-MeOH) extract and continuous ethanol (s-EtOH) extract pair obtained from the short modification of calcareous Chaetoceros The activity of Collagen of Fibroblasts albumen synthesis

Screen the continuous ethanol extraction of the direct methanol extract (d-MeOH) obtained from the short modification of calcareous Chaetoceros and two steps Thing (d-EtAc ＞ s-EtOH), to assess their adjustment effects to the synthesis of Collagen of Fibroblasts albumen.According to being previously described Experimental program extract is screened on fibroblast cell cultures.As a result it is reported in table 31.

Table 31

By handling primary fibroblast, the % ratios that control group is expressed as to the amount of activity-collagen of corium are detected

Embodiment	Sample	Quantity	It is average	Standard error	The quantity in hole	ANOVA
							0	Control	0	100.0	4.8	8	-
103	d-MeOH	0.1μg/ml	116.2	3.3	8	P ＜ 0.01
							104	d-MeOH	1.0μg/ml	113.2	2.7	8	P ＜ 0.01
105	d-MeOH	10μg/ml	118.0	4.2	8	P ＜ 0.01
							106	s-EtOH	0.01μg/ml	106.7	2.5	8	It is non-stimulated
107	s-EtOH	0.1μg/ml	112.7	3.5	8	P ＜ 0.01
							108	s-EtOH	1.0μg/ml	116.5	3.3	8	P ＜ 0.01

The synthesis of the extract increase collagen of two kinds of screenings.Cause optimal sound in the S-EtOH of 1.0 mcg/ml concentration Should, with 16% increase collagen synthesis, and the concentration of d-MeOH extracts induction optimal response is 10 mcg/mls, with The synthesis of 18% increase collagen.These results have the statistical significance (P ＜ 0.01) of highly significant.

Embodiment 109 to 114

Direct ethanol (d-EtOH) extract and continuous water (s- water) extract obtained from Thalassiosira is to fibroblast glue The activity of former albumen synthesis

The direct ethanol extract and continuous water that extraction of the screening by performing two steps is obtained from Thalassiosira pseudonana extract Thing, to assess their adjustment effects to the synthesis of Collagen of Fibroblasts albumen.According to previously described experimental program into fibre Extract is screened on dimension cell culture.As a result it is reported in table 32

Table 32

By handling primary fibroblast, the % ratios that control group is expressed as to the amount of activity-collagen of corium are detected

Embodiment	Sample	Quantity	It is average	Standard error	The quantity in hole	ANOVA
							0	Control	0	100.0	3.9	8	-
109	d-EtOH	0.1μg/ml	127.4	22.1	8	P ＜ 0.01
							110	d-EtOH	1.0μg/ml	148.9	6.0	8	P ＜ 0.01
111	d-EtOH	10μg/ml	148.4	5.4	8	P ＜ 0.01
							112	s-Water	0.1μg/ml	127.2	3.6	8	P ＜ 0.05
113	s-Water	1.0μg/ml	161.1	3.8	8	P ＜ 0.01
							114	s-Water	10μg/ml	172.5	10.1	8	P ＜ 0.01

From the extract that hailian seaweed obtains in all concentration for the treatment of to the synthetically produced strong impulse of collagen.It is described As a result there is the statistical significance of highly significant.

Embodiment 115 to 120

Direct ethanol (d-EtOH) extract and continuous water (s- water) extract obtained from Chlorococcum is to fibroblast glue The activity of former albumen synthesis

The direct ethanol extract and continuous water extract that extraction of the screening by performing two steps is obtained from Chlorococcum, with Assess their adjustment effects to the synthesis of Collagen of Fibroblasts albumen.According to previously described experimental program in fibroblast Extract is screened on culture.As a result it is reported in table 33.

Table 33

By handling primary fibroblast, the % ratios that control group is expressed as to the amount of activity-collagen of corium are detected

Embodiment	Sample	Quantity	It is average	Standard error	The quantity in hole	ANOVA
							0	Control	0	100.0	7.3	8	-
115	d-EtOH	0.1μg/ml	161.5	12.0	8	P ＜ 0.01
							116	d-EtOH	1.0μg/ml	153.0	12.4	8	P ＜ 0.01
117	d-EtOH	10μg/ml	186.9	8.8	8	P ＜ 0.01
							118	s-Water	0.1μg/ml	182.1	10.8	8	P ＜ 0.01
119	s-Water	1.0μg/ml	187.2	13.5	8	P ＜ 0.01
							120	s-Water	10μg/ml	150.6	10.8	8	P ＜ 0.01

From the extract that Chlorococcum obtains in all concentration for the treatment of to the synthetically produced strong impulse of collagen.Institute Stating result has the statistical significance of highly significant.

Embodiment 121 to 129

The activity that the continuous extract of three steps obtained from garlic Trentepohlia synthesizes Collagen of Fibroblasts albumen

According to previously described screening scheme, the continuous extract of three steps (the d-EtAc ＞ s-EtOH ＞ obtained from garlic algae are screened S- water) with assessment they to Collagen of Fibroblasts albumen synthesis adjustment effect.As a result it is reported in table 34：

Table 34

By handling primary fibroblast, the % ratios that control group is expressed as to the amount of activity-collagen of corium are detected

Embodiment	Sample	Quantity	It is average	Standard error	The quantity in hole	ANOVA
							0	Control	0	100	6.1	8	-
121	d-EtAc	0.1μg/ml	97.8	5.7	8	It is non-stimulated
							122	d-EtAc	1.0μg/ml	105.8	3.3	8	It is non-stimulated
123	d-EtAc	10μg/ml	110.5	5.0	8	It is non-stimulated
							124	s-EtOH	0.1μg/ml	106.4	5.4	8	It is non-stimulated
125	s-EtOH	1.0μg/ml	112.8	6.7	8	It is non-stimulated
							126	s-EtOH	10μg/ml	124.0	6.4	8	P ＜ 0.01
127	s-Water	0.1μg/ml	113.6	6.9	8	It is non-stimulated
							128	s-Water	1.0μg/ml	143.2	4.0	7	P ＜ 0.01
129	s-Water	10μg/ml	133.8	4.0	7	P ＜ 0.01

Synthetically produced thorn strongly of the continuous ethanol extract and continuous water extract obtained from garlic algae to collagen Swash.Cause the increase by 24% of the synthesis of collagen with the processing of the s-EtOH extracts of 1.0 mcg/mls, and it is micro- with 1.0 The processing of grams per milliliter and the s- water extracts of 10 mcg/mls, respectively with the synthesis of 43% and 33% increase collagen.Institute Stating result has the statistical significance of highly significant.Compared with the control group, d-EtAc extracts generate collagen synthesis The stimulation of appropriateness, up to 10%.

The adjusting of hyaluronic acid synthesis is studied by primary dermal fibroblast

According to following experimental program, screening analysis is carried out in the primary fibroblast culture handled with microalgae extract：

15,000 cells/wells of ■ are inoculated in 24 orifice plates and are cultivated in the complete medium of 500 microlitres/hole；

As long as ■ cells reach the convergence of about 80-90%, culture medium and being supplemented with microalgae extract with 500 microlitres/hole are removed Culture medium without FBS substitute.Control group receives the culture medium without FBS without supplement；

After the processing of 16 hours of ■, recycle culture medium and handle for hyaluronic acid ELISA test (Corgenix hyalomitomes Sour ELISA kit), and the plate comprising cellular layer is subjected to MTT tests, for cell quantity valuation；

To each groups of cells, the quantization of the hyaluronic acid of acquisition is standardized ■ in MTT data, and is expressed as relative to control group Performance percentage.

Embodiment 130 to 132

The activity synthesized from the extract that garlic Trentepohlia and Chlorococcum obtain to fibroblast hyaluronic acid

Using described experimental program, for being contrasted with the direct acetic acid ethyl acetate extract obtained from Chlorococcum (C), screening The direct ethyl acetate extract and continuous water extract obtained from garlic algae (M).In three experiments repeated, into fiber finer Born of the same parents' processing carries out under 1.0 mcg/mls.As a result it is reported in table 35.

Table 35

The % ratios of control group are expressed as to the amount of activity-hyaluronic acid of primary fibroblast

Embodiment	Sample	Strain	Quantity	It is average	Standard error	The quantity in hole	ANOVA
								0	Control	-	0	100.0	3.1	3
130	d-EtAc	C	1.0μg/ml	113.3	7.3	3	It is non-stimulated
								131	d-EtAc	M	1.0μg/ml	146.0	15.6	3	P ＜ 0.05
132	s-Water	M	1.0μg/ml	117.3	11.5	3	It is non-stimulated

Compared with the control group, the increase by 13% to 46% of the yield of fibroblastic hyaluronic acid of processing.By garlic algae The Activity Results that direct ethyl acetate extract represents have significant statistical significance.

Embodiment 133 to 134

Direct ethyl acetate (d-EtAc) extract and continuous ethanol (s-EtOH) obtained from the short modification of calcareous Chaetoceros carries The activity for taking thing to synthesize fibroblast hyaluronic acid

Be used to screening using described experimental program the direct ethyl acetate extract that obtains from the short modification of calcareous Chaetoceros and Continuous ethanol extract.In three repeated experiments fibroblast is handled in 10 mcg/mls.As a result it is reported in table 36 In.

Table 36

The % ratios of control group are expressed as to the quantity of activity-hyaluronic acid of primary fibroblast

Embodiment	Sample	Quantity	It is average	Standard error	The quantity in hole	ANOVA
							0	Control	0	100	4.37	3
133	d-EtAc	10μg/ml	117.9	2.36	3	P ＜ 0.05
							134	s-EtOH	10μg/ml	106.8	9.1	3	It is non-stimulated

Compared with the control group, 18% is increased with the yield of fibroblastic hyaluronic acid of EtAc extract-treateds.By angle hair The Activity Results that the direct ethyl acetate extract of algae represents have statistical significance (P ＜ 0.05).

The adjusting of hyaluronic acid synthesis is studied by primary culture keratinocytes

According to following experimental program, screening analysis is carried out in the primary culture keratinocytes culture handled with microalgae extract：

25,000 cells/wells of ■ are inoculated in 24 orifice plates and are cultivated in the complete medium of 500 microlitres/hole；

As long as convergence of the ■ cells more than 50%, culture medium is removed and with 500 microlitres/hole with the culture of microalgae extract supplement Base substitutes.Control group receives the standard medium without supplement；

After the processing of 16 hours of ■, recycle culture medium and handle for hyaluronic acid ELISA test (Corgenix hyalomitomes Sour ELISA kit), and remaining cellular layer is subjected to MTT tests, for cell quantity valuation；

■ is to each groups of cells, and the quantization of the hyaluronic acid of acquisition is standardized in MTT data, and table

It is shown as the percentage of the performance relative to control group.

Embodiment 135 to 136

The activity of the extract Human Keratinocytes hyaluronic acid synthesis obtained from garlic Trentepohlia and Chlorococcum

Using described experimental program, compared with the direct ethyl acetate extract with being obtained from Chlorococcum (C), screening The direct ethyl acetate extract obtained from garlic algae (M).Keratinocyte is handled in 1.0 mcg/mls, repeats experiment 3 It is secondary.As a result it is reported in table 37.

Table 37

The % ratios of control group are expressed as to the amount of activity-hyaluronic acid of primary culture keratinocytes

Embodiment	Sample	Strain	Quantity	It is average	Standard error	The quantity in hole	ANOVA
								0	Control	-	0	100.0	1.4	3	-
135	d-EtAc	C	1.0μg/ml	102.9	0.9	3	It is non-stimulated
								136	d-EtAc	M	1.0μg/ml	111.3	4.0	3	P ＜ 0.05

The direct ethyl acetate extract obtained from garlic algae stimulates fibroblastic collagen synthesis increase by 11%.Response With statistical significance (P ＜ 0.05).

Conclusion

Above-mentioned experiment discloses the extract obtained from garlic algae, the short modification of calcareous Chaetoceros and Chlorococcum, by skin into fibre There is bioactivity in the synthesis for the hyaluronic acid that dimension cell and keratinocyte carry out.This strongly suggests that these are extracted Thing can be as the active ingredient for anti-aging and anti-photoaging formulation.

E. activity of the disclosed microalgae in vitro corium and epidermis

Number reported below is it was demonstrated that the microalgae extract of screening can be adjusted by fibroblast and keratinocyte progress Basic protein and GAG components synthesis.However, by being tested in vitro Full thickness human skin, we are into one Step have studied the property of microalgae.Following experiment purpose is：

By studying the collagen quantity in histotomy, confirm the synthesis of relevant collagen in isolated skin stimulates ■ Effect；

■ studies presence of the involucrin in cuticula by Western blot analysis (WB), and exploring microalgae extract can The differentiation of keratinocyte is adjusted in the epidermis of skin.

For the composition of corium, the dependent interaction of collagen had been discussed.The cuticula of skin is the surface portion of tissue, It is made of dead angle cell plastid, it completes related function：Prevent the moisture loss from skin and the organism from external environment condition Or the infiltration of undesirable compound.The integrality and function of cuticula are essential, to keep appropriate moisture of skin And safeguard invasion and attack of the body from pathogen.Skin needs moisture, to keep smooth and soft, but as body ages retain Moisture becomes more and more difficult.On the other hand, the change of the thickness and component of cuticula is some common skin disease (examples Such as, xerosis) or really pathologic conditions (i.e. psoriasis) the reason for or one of reason.The cuticle thickness of reduction may cause Skin dehydration, and in contrast the problem of may result in hyperkeratosis.The cell constituent of cuticula is horn cell, Horn cell represents the final differential period of epidermal keratinocytes.Involucrin is the angle based on Keratinocyte differentiation The protein labeling for the identification that change process is related to.The synthesis of involucrin can be studied, with define whether experimentally processing increase or Reduce epidermal cornified.These activity all have interests for the application in cosmetics and acology.

The synthesis of garlic algae and the continuous water extract of Chlorococcum to skin corium collagen is studied on excised human skin's culture Activity

Embodiment 137 to 140

Activity from the continuous extract that garlic Trentepohlia and Chlorococcum obtain to collagen synthesis in cultured in vitro skin

Continuously extracted by three steps from garlic algae (M) and Chlorococcum (C) cell material and prepare continuous water extract.1 microgram/ Milliliter and two kinds of different ultimate densities of 10 mcg/mls, these extracts are dissolved in pure DMSO.Relative to cultured in vitro The adjusting of the melanogenesis of skin, the organ for the skin for preparing through thickness people reported respectively such as (such as embodiment 58 to 63) Culture.These microalgae preparations being diluted in DMSO that the dermatological specimens of culture locally apply 4 microlitres daily are handled.Through Cross after the organ culture of six days, prepare histotomy from dermatological specimens, and fixed group is handled according to Miller staining techniques Fabric products, study the quantitative change of collagen.The microphoto of capturing skin histotomy, by submitting microphoto to electricity Brain image analysis obtains dermal collagen and quantifies.The results are shown in table 38.

Table 38

Conjunction of the continuous water extract obtained from garlic Trentepohlia and Chlorococcum to the collagen in the application on human skin of cultured in vitro Into activity-collagen amount be expressed as control group performance % ratios

Embodiment	Sample	Strain	Quantity	It is average	Standard error	Sample size	ANOVA
								0	Control	-	0	100.0	10.3	12	-
137	s-Water	C	1.0μg/ml	121.5	7.1	12	It is non-stimulated
								138	s-Water	C	10μg/ml	118.5	11.8	12	It is non-stimulated
139	s-Water	M	1.0μg/ml	143.8	13.1	12	P ＜ 0.01
								140	s-Water	M	10μg/ml	143.6	9.4	11	P ＜ 0.01

The results show that the synthesis of all continuous water extract stimulation collagens, the response excursion is 18% to 44%. Statistical significance of the response with highly significant produced with the processing of garlic Trentepohlia s- water extracts.

Embodiment 141 to 152

The activity synthesized from the continuous extract that garlic Trentepohlia and Chlorococcum obtain to the involucrin of cultured in vitro skin

Using previously described skin organ culture technique, for screening from the continuous of garlic algae (M) and Chlorococcum (C) acquisition Extract, to explore their activity to horny layer of epidermis.It is 1 micro- by dissolving dry extracts to reach ultimate density in DMSO Grams per milliliter and 10 mcg/mls, prepare Local treatment.The dermatological specimens for handling culture with these preparations daily continue six days, so Analyzed afterwards by Western blot and quantify involucrin content.Preparing protein extract it is analyzed for Western blot Before, corium is almost removed from skin samples.Each experimental group is handled, merges 3 in the preparation process of protein extract A dermatological specimens, then each protein extract pass through Western blot technologies carry out replicate analysis.The sound of the tissue of processing It should be reported in table 39, the change percentage for the involucrin content being expressed as compared with control group.

Table 39

What the involucrin in application on human skin of the continuous water extract obtained from Chlorococcum and garlic algae to cultured in vitro synthesized The quantity of activity-involucrin is expressed as the % ratios of the performance of control group

Compared with control group, the extract that the slave Chlorococcum of test obtains stimulates the synthesis of involucrin to increase by 10% to 75%, And continuous water extract causes the response (75%) of higher.

The direct ethyl acetate obtained from garlic algae and continuous ethanol extract stimulate the synthesis increase by 22% of involucrin to 34%, and continuous water extract reduces involucrin synthesis.

F. for the activity of cell proliferation disclosed in the microalgae extract of consideration

Skin cell proliferation represents relevant issues at least two reasons：The holding and wound healing of melanocyte.

Normal human melanocytes are located at the basalis of epidermis, seldom carry out mitosis.When body aging in some epidermises (such as, for example, in hair follicle) their density can be reduced in structure.Therefore, cosmetic industry is interested in suitable very much Promote the activity of melanocyte and the product of propagation.In addition, some skin diseases, such as leucoderma, are due to melanocyte Posteriority depigmentation caused by death, therefore, active compound may be for similar in melanocyte proliferation The treatment of problem is effective.

Meanwhile when producing wound in the tissue, the propagation of other Skin Cells is desirably.Increase the propagation of cell Rate, reduces the risk of infection, accelerating wound healing process.Treatment for unexpected injury and skin disease or beauty treatment, it is necessary to Support the product of the process of wound healing.

Some extracts obtained from above-mentioned microalgae are screened, to assess the effect of their cell proliferations.Make us frightened It is surprised, the relevant stimulation of suitable concentration their cell proliferation rates generation ought be employed by disclosing, and in the thin of sensitivity Born of the same parents' species produces relevant stimulation.

Embodiment 153 to 160

Direct methanol (d-MeOH) extract obtained from Chaetoceros category, Thalassiosira, garlic Trentepohlia and Chlorococcum is to black The proliferation activity of plain class cell

Experiment is intended to research and obtains from Chaetoceros category (K), Thalassiosira (T), garlic Trentepohlia (M) and Chlorococcum (C) below The effect of direct methanol (d-MeOH) extract of acquisition.

B16V cells are chosen to be the reactive representative model of melanocyte.96 orifice plates are inoculated with 7500 cells/wells, and carefully Born of the same parents are cultivated in the culture medium RPMI 1640 supplemented with 10%FBS and 4mM glutamine.Cultivate 24 it is small when after, verification The health status of cell, and the culture medium is substituted with start to process with the culture medium of experiment.Repeated in six holes per treatment.It is right In the extract of each screening, by respectively be dissolved in DMSO the 20,000 of 0.5 microlitre/milliliter, or 0.05 microlitre/milliliter The dry microalgae extract supplement standard medium of mcg/ml prepares the assay medium.In the culture medium of these supplements Final extract concentrations are respectively 10 mcg/mls, 0.1 mcg/ml, and DMSO concentration is 0.05% and 0.005%.Two Control group is handled by the standard medium supplemented with 0.05%, 0.005%DMSO respectively.After the processing of 3 days, stop Only cultivate, the final cell quantity of corresponding each experimental group passes through resazurin test assessment.

As a result the average percent change for the cell density being expressed as compared with control group, and it is comprehensive in following table 40.

Table 40

Pass through the work of the d-MeOH extract-treateds B16V of the 0.1 mcg/ml and 10 mcg/mls cell proliferations detected The final amt of property-cell is expressed as the % ratios of control group performance

As a result prove that this is stimulated carefully from the extract that Chaetoceros, hailian seaweed and Chlorococcum obtain when concentration is 10 mcg/ml Born of the same parents breed, and have the statistical significance of statistical significance or highly significant.

Embodiment 161 to 169

Activity from the continuous extract of three steps that Chlorococcum obtains to fibroblast proliferation

Experimental arrangement is based on following steps：

■ primary fibroblasts are inoculated in 96 hole microwell plates, density for 20000 cells/centimetre²；

After when ■ cultures 24 are small, culture medium is replaced with the culture medium through supplement, the culture medium of supplement by add 0.1 microgram/ It is prepared by the extract that the slave Chlorococcum cell material of milliliter, 1.0 mcg/mls or 10 mcg/mls obtains.Octal dedicated for The screening of the culture medium each supplemented.Keep octal as a control group, and cultivated in standard medium；

After when culture 48-72 is small, fibroblast density is assessed by MTT experiment, and the performance of each experimental group is expressed For the average percent of the performance of control group.The results are shown in table 41：

Table 41

0.1st, the continuous extract-treated primary fibroblast that the slave Chlorococcum of 1 and 10 mcg/mls obtains detects The final amt of activity-cell of cell proliferation is expressed as the % ratios of control group performance

Embodiment	Sample	Quantity	It is average	Standard error	The quantity in hole	ANOVA
							0	Control	0	100.0	2.8	8	-
161	d-EtAc	0.1μg/ml	105.9	7.9	8	It is non-stimulated
							162	d-EtAc	1.0μg/ml	120.3	2.6	8	P ＜ 0.01
163	d-EtAc	10μg/ml	124.8	4.4	8	P ＜ 0.01
							164	s-EtOH	0.1μg/ml	109.4	5.4	8	It is non-stimulated
165	s-EtOH	1.0μg/ml	113.3	2.6	8	P ＜ 0.05
							166	s-EtOH	10μg/ml	93.1	5.3	8	It is non-stimulated
167	s-Water	0.1μg/ml	117.4	4.7	8	P ＜ 0.01
							168	s-Water	1.0μg/ml	114.2	3.4	8	P ＜ 0.05
169	s-Water	10μg/ml	100.3	5.1	8	It is non-stimulated

If these numbers using appropriate all continuous extracts of concentration it was demonstrated that can be stimulated cellular proliferation.With control group phase Than the processing makes final cell quantity increase by 13% to 25%, has statistical significance.

Embodiment 170 to 178

Activity from the continuous extract of three steps that garlic Trentepohlia obtains to fibroblast proliferation

The continuous extract that the cell material that identical experimental arrangement is used to screen from garlic algae obtains.The results are shown in table 42 In：

Table 42

0.1st, pair that the continuous extract-treated primary fibroblast that the slave garlic algae of 1 and 10 mcg/mls obtains detects The final amt of activity-cell of cell Proliferation is expressed as the % ratios of control group performance

These numbers are it was demonstrated that compared with control group, in all cases, in addition to the processing of 0.1 mcg/ml dETAC, institute There is continuous extract to stimulate cellular proliferation, cause final cell quantity to increase by 9% to 27%, be respectively provided with statistical significance.

G. the activity for being decomposed disclosed in the microalgae extract that is considered to people's adipocyte fatty

In order to assess activity of the above-mentioned microalgae to lipid-metabolism, cultivate and with from Chaetoceros category, Thalassiosira, garlic Trentepohlia and green The d-MeOH extract-treated application on human skin samples that ball Trentepohlia obtains.

With untreated group of contrast, the reaction of the tissue of processing, is commented by measuring the dissociative glycerin discharged in the medium Valency, the primary final products of dissociative glycerin, that is, lipid-metabolism.

Embodiment 179 to 186

The d-MeOH extracts obtained from Chaetoceros category, Thalassiosira, garlic Trentepohlia and Chlorococcum are to full thickness cutaneous culture The activity of thing

Carried using the direct methanol obtained from Chaetoceros category (K), Thalassiosira pseudonana (T), garlic algae (M) and Chlorococcum (C) Thing is taken to be used for the sample for supplementing modified William's culture medium E, for cultivating the application on human skin of through thickness.It is every kind of micro- in supplementing culture medium The ultimate density of algae extract is respectively 5 and 50 mcg/mls.

In order to make activity of the assessment microalgae extract to skin lipid metabolism, using following experimental procedure：

■ cuts off the cylindrical application on human skin sample of 7 millimeters of diameter from same donor respectively, and is inoculated in the density in 1 sample/hole 24 orifice plates of the standard medium with 700 microlitres/hole；

After culture when ■ 24 is small, standard medium is replaced with assay medium, and in addition to control group, control group is for the first time Receive standard medium.Each experiment process gives 4 dermatological specimens (in four different holes).Control group also includes four samples Product.

After processing when ■ 24 is small, quantify cultivating by using the dissociative glycerin reagent (code #F6428) of Sigma productions The glycerine discharged in base.25 microlitres of culture medium is taken from each hole and divides three parts and is put into 96 orifice plates, is separately added into 200 microlitres Dissociative glycerin reagent into each hole.It is incubated at room temperature after 20 minutes, plate microplate luminometer is read at 540nm Take.The standard curve of the glycerine of (relying) as reference, quantification culture medium are depended on by the absorbance value for making to detect In dissociative glycerin.

Experimental result is expressed as the percentage of the performance of control group, and the results are shown in table 43.

Table 43

The skin fat that the d-MeOH extracts obtained from Chaetoceros category, Thalassiosira pseudonana, garlic algae and Chlorococcum carry out Decompose the % ratios that adjusting-data are represented as the performance of control group

Embodiment	Sample	Quantity	It is average	Standard error	The quantity in hole	ANOVA
							0	Control	0	100.0	14.2	4
179	K	5.0μg/ml	136.9	10.1	4	P ＜ 0.05
							180	K	50μg/ml	99.3	7.6	4	It is non-stimulated
181	T	5.0μg/ml	147.9	9.4	4	P ＜ 0.01
							182	T	50μg/ml	109.3	16.8	4	It is non-stimulated
183	M	5.0μg/ml	141.1	3.8	4	P ＜ 0.05
							184	M	50μg/ml	123.7	16.3	4	It is non-stimulated
185	C	5.0μg/ml	142.4	1.5	4	P ＜ 0.05
							186	C	50μg/ml	152.9	14.3	4	P ＜ 0.01

If experimental result confirms that all extracts stimulate lipolysis to act on using suitable concentration, compared with control group, The release increase by 23% to 53% of dissociative glycerin.

The extract obtained from diatom (Chaetoceros and hailian seaweed) has high dry weight (being shown in Table 1), induces and has in 5 mcg/mls The response of statistically significant or highly significant statistical significance, and they do not produce any shadow in 50 mcg/ml Ring.

On the contrary, skin fat is stimulated to decompose in all concentration for the treatment of with the processing that garlic algae and Chlorococcum extract carry out, to the greatest extent Pipe does not have statistical significance by using the response that the garlic Trentepohlia extract-treated skin detection of 50 mcg/mls arrives.

The microalgae extract of screening discloses the lipolysis that can adjust skin strongly.They are suitable for being included in for beauty In the preparation for being used to treat adipose tissue disease of therapeutic purposes.

Embodiment 187 to 194

The d-MeOH extracts obtained from Chaetoceros category, Thalassiosira, garlic Trentepohlia and Chlorococcum are to full thickness cutaneous culture The activity of thing

The experimental procedure of experiment description above is repeated, to confirm the bioactivity of the methanol preparation obtained from the microalgae of consideration. Obtained data are shown in table 44.

Table 44

The skin fat that the d-MeOH extracts obtained from Chaetoceros category, Thalassiosira pseudonana, garlic algae and Chlorococcum carry out Decompose the % ratios that adjusting-data are represented as the performance of control group

Embodiment	Sample	Quantity	It is average	Standard error	The quantity in hole	ANOVA
							0	Control	0	100.0	15.7	4
187	K	5.0μg/ml	192.0	15.8	4	P ＜ 0.01
							188	K	50μg/ml	104.3	10.0	4	It is non-stimulated
189	T	5.0μg/ml	109.3	17.5	4	It is non-stimulated
							190	T	50μg/ml	102.0	23.5	4	It is non-stimulated
191	M	5.0μg/ml	195.3	24.3	4	P ＜ 0.01
							192	M	50μg/ml	173.4	10.3	4	P ＜ 0.01
193	C	5.0μg/ml	214.8	11.0	4	P ＜ 0.01
							194	C	50μg/ml	202.2	13.9	4	P ＜ 0.01

The bioactivity of the stimulation lipolysis of data confirm that extract, has strong especially for garlic algae and Chlorococcum extract Bioactivity, it adds the release of dissociative glycerin in all concentration for the treatment of.Compared with control group, both extraction produce The stimulation of raw lipolysis increases by 73% to 114%, always the statistical significance with highly significant.

The extract obtained from diatom is confirmed as more active in the case where 5 mcg/mls are than the concentration in higher.Although two kinds of systems Agent stimulates lipolysis, but only Chaetoceros have statistical significance.

Embodiment 195 to 198

Activity from the d-MeOH extracts that Thalassiosira and Chlorococcum obtain to full thickness cutaneous culture

The experimental procedure of experiment description above is repeated, to confirm the methanol preparation obtained from Thalassiosira pseudonana and Chlorococcum Bioactivity.Obtained data are shown in table 45.

Table 45

The skin fat that the d-MeOH extracts obtained from Thalassiosira pseudonana and Chaetoceros category carry out decomposes adjusting-data by table It is shown as the % ratios of the performance of control group

Embodiment	Sample	Quantity	It is average	Standard error	The quantity in hole	ANOVA
							0	Control	0	100.0	10.9	4
195	T	5.0μg/ml	127.0	25.5	4	It is non-stimulated
							196	T	50μg/ml	140.4	10.0	4	It is non-stimulated
197	C	5.0μg/ml	116.6	30.1	4	It is non-stimulated
							198	C	50μg/ml	169.8	24.5	4	P ＜ 0.01

Data confirm that extract stimulates the bioactivity of lipolysis.In response to the processing with hailian seaweed extract, fat point The peak value for increasing up to 40% of solution, and in response to the processing with Chlorococcum extract, it increases by 70%.

Embodiment 199 to 206

From the d-MeOH extracts that Chaetoceros category, the short modification of calcareous Chaetoceros, Chlorococcum and small Chlorococcum obtain to complete thick Spend the activity of Skins culture thing

The experimental procedure of experiment description above is repeated, with the bioactivity of the different algae strains of more same category.From calcareous angle The short modification of hair algae (also known as CCAP 1010/11 and algae and the protozoan culture for being preserved in the management of Scotland Marine Sciences association Thing collection) dry biomass and small Chlorococcum (also referred to as CCAP 213/7 simultaneously be stored in algae and protozoan training Yang Wu collecting center) dry biomass prepare new methanolic extract.Using these new extracts with from Chaetoceros category and from The previous preparation that Chlorococcum (both source is unknown) obtains compares, for handling the people of people's full thickness cutaneous treatment holostrome Dermatological specimens, to study their effects to lipolysis.Obtained data are shown in table 46.

Table 46

From the short modification of calcareous Chaetoceros (K-CCAP1010/11), Chaetoceros category (K), small Chlorococcum (C-CCAP213/7) and green The skin fat that the d-MeOH extracts that ball Trentepohlia (C) obtains carry out decomposes the performance that adjusting-data are represented as control group % ratios

Embodiment	Sample	Quantity	It is average	Standard error	The quantity in hole	ANOVA
							0	Control	0	100.0	12.5	4
199	K	5.0μg/ml	118.7	11.6	4	It is non-stimulated
							200	K	50μg/ml	115.5	7.6	4	It is non-stimulated
201	K ccap1010/11	5.0μg/ml	136.2	3.8	4	P ＜ 0.05
							202	K ccap1010/11	50μg/ml	109.6	10.3	4	It is non-stimulated
203	C	5.0μg/ml	139.9	8.8	4	P ＜ 0.05
							204	C	50μg/ml	117.1	3.0	4	It is non-stimulated
205	Cccap213/7	5.0μg/ml	123.2	12.9	4	It is non-stimulated
							206	Cccap213/7	50μg/ml	118.1	17.5	4	It is non-stimulated

The processing makes the release of dissociative glycerin add 9% to 40%, it was confirmed that the thorn of the lipolysis of the algae strain of same category Swash effect.However, in the case of the preparation obtained from Chlorococcum and the short modification CCAP1010/11 of calcareous Chaetoceros, it is described Preparation is more active preparation, it produces the response with statistical significance.

Claims (10)

1. cosmetic composition, it includes garlic Trentepohlia and/or the extract and U.S. of Chaetoceros category and/or Chlorococcum microalgae Acceptable auxiliary material in appearance, the cosmetically acceptable auxiliary material are selected from C₁-C₄Aliphatic alcohol, have the polynary of 3 to 12 carbon atoms Alcohol, oil ingredient, water and their mixture, wherein the extract is by using selected from C₁-C₄Aliphatic alcohol, ethyl acetate, water or it The solvent of mixture handle the microalgae, the extract of dissolving is removed from residue and from the pure institute of the solvent recovery Extract is stated to obtain.

2. composition according to claim 1 is used for the purposes for treating application on human skin.

3. composition according to claim 1 is used for the purposes for treating people's hair.

4. composition according to claim 1 is used for the purposes for suppressing the unnecessary growth of people's hair.

5. composition according to claim 1 is used for the purposes for adjusting the melanogenesis in people's hair and/or application on human skin.

6. composition according to claim 1 is used for the purposes for the growth for improving and stimulating people's hair and hair follicle.

7. composition confrontation according to claim 1 and the purposes of prevention trichomadesis.

8. composition according to claim 1 is used to improve and stimulates the purposes of the collagen synthesis in human dermis' layer.

9. composition according to claim 1 is used to prevent and the purposes to resisting age of skin.

10. composition according to claim 1 is used to improving and stimulating the purposes that the glucosaminoglycan in application on human skin synthesizes.